Effects of a spaceflight environment on heritable changes in wheat gene expression.

Once it was established that the spaceflight environment was not a drastic impediment to plant growth, a remaining space biology question was whether long-term spaceflight exposure could cause changes in subsequent generations, even if they were returned to a normal Earth environment. In this study, we used a genomic approach to address this question. We tested whether changes in gene expression patterns occur in wheat plants that are several generations removed from growth in space, compared to wheat plants with no spaceflight exposure in their lineage. Wheat flown on Mir for 167 days in 1991 formed viable seeds back on Earth. These seeds were grown on the ground for three additional generations. Gene expression of fourth-generation Mir flight leaves was compared to that of the control leaves by using custom-made wheat microarrays. The data were evaluated using analysis of variance, and transcript abundance of each gene was contrasted among samples with t-tests. After corrections were made for multiple tests, none of the wheat genes represented on the microarrays showed a statistically significant difference in expression between wheat that has spaceflight exposure in their lineage and plants with no spaceflight exposure. This suggests that exposure to the spaceflight environment in low Earth orbit space stations does not cause significant, heritable changes in gene expression patterns in plants.

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.

Technical review of the Laboratory Biosphere closed ecological system facility.

Laboratory Biosphere is a 40 m3 closed life system that commenced operation in May 2002. Light is from 12,000 W of high pressure sodium lamps over planting beds with 5.37 m2 of soil. Water is 100% recycled by collecting condensate from the temperature and humidity control system and mixing with leachate collected from under the planting beds. Atmospheric leakage was estimated during the first closure experiment to be 0.5-1% per day in general plus about 1% for each usage of the airlock door. The first trial run of 94 days was with a soybean crop grown from seeds (May 17, 2002) to harvest (August 14, 2002) plus 5 days of post-harvest closure. The focus of this initial trial was system testing to confirm functionality and identify any necessary modifications or improvements. This paper describes the organizational and physical features of the Laboratory Biosphere.

Earth applications of closed ecological systems: relevance to the development of sustainability in our global biosphere.

The parallels between the challenges facing bioregenerative life support in artificial closed ecological systems and those in our global biosphere are striking. At the scale of the current global technosphere and expanding human population, it is increasingly obvious that the biosphere can no longer safely buffer and absorb technogenic and anthropogenic pollutants. The loss of biodiversity, reliance on non-renewable natural resources, and conversion of once wild ecosystems for human use with attendant desertification/soil erosion, has led to a shift of consciousness and the widespread call for sustainability of human activities. For researchers working on bioregenerative life support in closed systems, the small volumes and faster cycling times than in the Earth's biosphere make it starkly clear that systems must be designed to ensure renewal of water and atmosphere, nutrient recycling, production of healthy food, and safe environmental methods of maintaining technical systems. The development of technical systems that can be fully integrated and supportive of living systems is a harbinger of new perspectives as well as technologies in the global environment. In addition, closed system bioregenerative life support offers opportunities for public education and consciousness changing of how to live with our global biosphere.

Initial experimental results from the Laboratory Biosphere closed ecological system facility.

An initial experiment in the Laboratory Biosphere facility, Santa Fe, New Mexico, was conducted May-August 2002 using a soil-based system with light levels (at 12 h per day) of 58-mol m-2 d-1. The crop tested was soybean, cultivar Hoyt, which produced an aboveground biomass of 2510 grams. Dynamics of a number of trace gases showed that methane, nitrous oxide, carbon monoxide, and hydrogen gas had initial increases that were substantially reduced in concentration by the end of the experiment. Methane was reduced from 209 ppm to 11 ppm, and nitrous oxide from 5 ppm to 1.4 ppm in the last 40 days of the closure experiment. Ethylene was at elevated levels compared to ambient during the flowering/fruiting phase of the crop. Soil respiration from the 5.37 m2 (1.46 m3) soil component was estimated at 23.4 ppm h-1 or 1.28 g CO2 h-1 or 5.7 g CO2 m-2 d-1. Phytorespiration peaked near the time of fruiting at about 160 ppm h-1. At the height of plant growth, photosynthesis CO2 draw down was as high as 3950 ppm d-1, and averaged 265 ppm h-1 (whole day averages) during lighted hours with a range of 156-390 ppm h-1. During this period, the chamber required injections of CO2 to continue plant growth. Oxygen levels rose along with the injections of carbon dioxide. Upon several occasions, CO2 was allowed to be drawn down to severely limiting levels, bottoming at around 150 ppm. A strong positive correlation (about 0.05 ppm h-1 ppm-1 with r2 about 0.9 for the range 1000-5000 ppm) was observed between atmospheric CO2 concentration and the rate of fixation up to concentrations of around 8800 ppm CO2.

Advantages of using subsurface flow constructed wetlands for wastewater treatment in space applications: ground-based Mars Base prototype.

Research and design of subsurface flow wetland wastewater treatment systems for a ground-based experimental prototype Mars Base facility has been carried out, using a subsurface flow approach. These systems have distinct advantages in planetary exploration scenarios: they are odorless, relatively low-labor and low-energy, assist in purification of water and recycling of atmospheric CO2, and will support some food crops. An area of 6-8 m2 may be sufficient for integration of wetland wastewater treatment with a prototype Mars Base supporting 4-5 people. Discharge water from the wetland system will be used as irrigation water for the agricultural crop area, thus ensuring complete recycling and utilization of nutrients. Since the primary requirements for wetland treatment systems are warm temperatures and lighting, such bioregenerative systems may be integrated into early Mars base habitats, since waste heat from the lights may be used for temperature maintenance in the human living environment. "Wastewater gardens (TM)" can be modified for space habitats to lower space and mass requirements. Many of its construction requirements can eventually be met with use of in-situ materials, such as gravel from the Mars surface. Because the technology requires little machinery and no chemicals, and relies more on natural ecological mechanisms (microbial and plant metabolism), maintenance requirements are minimized, and systems can be expected to have long operating lifetimes. Research needs include suitability of Martian soil and gravel for wetland systems, system sealing and liner options in a Mars Base, and wetland water quality efficiency under varying temperature and light regimes.

Potential integration of wetland wastewater treatment with space life support systems.

Subsurface-flow constructed wetlands for wastewater treatment and nutrient recycling have a number of advantages in planetary exploration scenarios: they are odorless, relatively low labor and low energy, assist in purification of water and recycling of atmospheric CO2, and can directly grow some food crops. This article presents calculations for integration of wetland wastewater treatment with a prototype ground-based experimental facility ("Mars on Earth") supporting four people showing that an area of 4-6 m2 may be sufficient to accomplish wastewater treatment and recycling. Discharge water from the wetland system can be used as irrigation water for the agricultural crop area, thus ensuring complete reclamation and utilization of nutrients within the bioregenerative life support system. Because the primary requirements for wetland treatment systems are warm temperatures and lighting, such bioregenerative systems can be integrated into space life support systems because heat from the lights may be used for temperature maintenance in the human living environment. Subsurface-flow wetlands can be modified for space habitats to lower space and mass requirements. Many of its construction requirements can eventually be met with use of in situ materials, such as gravel from the Mars surface. Because the technology does not depend on machinery and chemicals, and relies more on natural ecological mechanisms (microbial and plant metabolism), maintenance requirements (e.g., pumps, aerators, and chemicals) are minimized, and systems may have long operating lifetimes. Research needs include suitability of Martian soil and gravel for wetland systems, system sealing and liner options in a Mars base, and determination of wetland water quality efficiency under varying temperature and light regimes.

Light, plants, and power for life support on Mars.

Regardless of how well other growing conditions are optimized, crop yields will be limited by the available light up to saturation irradiances. Considering the various factors of clouds on Earth, dust storms on Mars, thickness of atmosphere, and relative orbits, there is roughly 2/3 as much light averaged annually on Mars as on Earth. On Mars, however, crops must be grown under controlled conditions (greenhouse or growth rooms). Because there presently exists no material that can safely be pressurized, insulated, and resist hazards of puncture and deterioration to create life support systems on Mars while allowing for sufficient natural light penetration as well, artificial light will have to be supplied. If high irradiance is provided for long daily photoperiods, the growing area can be reduced by a factor of 3-4 relative to the most efficient irradiance for cereal crops such as wheat and rice, and perhaps for some other crops. Only a small penalty in required energy will be incurred by such optimization. To obtain maximum yields, crops must be chosen that can utilize high irradiances. Factors that increase ability to convert high light into increased productivity include canopy architecture, high-yield index (harvest index), and long-day or day-neutral flowering and tuberization responses. Prototype life support systems such as Bios-3 in Siberia or the Mars on Earth Project need to be undertaken to test and further refine systems and parameters.

Engineering of Biosphere 2: closure and energy.

As an experimental apparatus, Biosphere 2 was predicated on material closure to permit investigation of ecological processes without unknown sources or sinks outside the ecosystems under study. The atmospheric leak rate is demonstrated to be less than 10% per year. The enabling technology to achieve the low leak rate is expansion chambers called "lungs." Closure enables use of mass balance equations to track chemical pathways (e.g., of carbon dioxide and oxygen). Energy and information pass across the enclosure without incurring any material transfer. Water is fully recycled.

Living in space: results from Biosphere 2's initial closure, an early testbed for closed ecological systems on Mars.

The following summary of results from the first 2-year closure experiment (September 26, 1991 to September 26, 1993) in Biosphere 2 is excerpted from a chapter written by William Dempster and myself for a book, Strategies for Mars, edited by Carol Stoker and Carter Emmart of NASA Ames Research Center. The book will be published later this year by Krieger Publishers. It brings together a number of the most striking initial results, including food production and nutrition; ecosystem changes; oxygen and carbon dioxide dynamics; and the human role and response to living in a small, recycling life support system. The references cited are useful as a guide to currently available articles in journals. Hopefully, the next year will see a proliferation of papers presenting more data from the first 2 years of Biosphere 2's operation. There was a wealth of data collected during the closure and by teams of researchers who had access to the facility during the 5-month transition period following the departure of the first crew and the commencement of the second closure experiment in March, 1994.

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

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

Biosphere 2: a prototype project for a permanent and evolving life system for Mars base.

As part of the ground-based preparation for creating long-term life systems needed for space habitation and settlement, Space Biospheres Ventures (SBV) is undertaking the Biosphere 2 project near Oracle, Arizona. Biosphere 2, currently under construction, is scheduled to commence its operations in 1991 with a two-year closure period with a crew of eight people. Biosphere 2 is a facility which will be essentialy materially-closed to exchange with the outside environment. It is open to information and energy flow. Biosphere 2 is designed to achieve a complex life-support system by the integration of seven areas or "biomes"--rainforest, savannah, desert, marsh, ocean, intensive agriculture and human habitat. Unique bioregenerative technologies, such as soil bed reactors for air purification, aquatic waste processing systems, real-time analytic systems and complex computer monitoring and control systems are being developed for the Biosphere 2 project. Its operation should afford valuable insight into the functioning of complex life systems necessary for long-term habitation in space. It will serve as an experimental ground-based prototype and testbed for the stable, permanent life systems needed for human exploration of Mars.

Biosphere II: engineering of manned, closed ecological systems.

Space Biospheres and Ventures, a private, for-profit firm, has undertaken a major research and development project in the study of biospheres, with the objective of creating and producing biospheres. Biosphere II-scheduled for completion in March 1991-will be essentially isolated from the existing biosphere by a closed structure, composed of components derived from the existing biosphere. Like the biosphere of the Earth, Biosphere II will be essentially closed to exchanges of material or living organisms with the surrounding environment and open to energy and information exchanges. Also, like the biosphere of the Earth, Biosphere II will contain five kingdoms of life, a variety of ecosystems, plus humankind, culture, and technics. The system is designed to be complex, stable and evolving throughout its intended 100-year lifespan, rather than static. Biosphere II will cover approximately 1.3 hectare and contain 200,000 m3 in volume, with seven major biomes: tropical rainforest, tropical savannah, marsh, marine, desert, intensive agriculture, and human habitat. An interdisciplinary team of leading scientific, ecological, management, architectural, and engineering consultants have been contracted by Space Biospheres Ventures for the project. Potential applications for biospheric systems include scientific and ecological management research, refuges for endangered species, and life habitats for manned stations on spacecraft or other planets.