Understanding plant-soil relationships using controlled environment facilities.

Although soil is a component of terrestrial ecosystems, it is comprised of a complex web of interacting organisms, and therefore can be considered itself as an ecosystem. Soil microflora and fauna derive energy from plants and plant residues and serve important functions in maintaining soil physical and chemical properties, thereby affecting net primary productivity (NPP), and in the case of contained environments, the quality of the life support system. We have been using 3 controlled-environment facilities (CEF's) that incorporate different levels of soil biological complexity and environmental control, and differ in their resemblance to natural ecosystems, to study relationships among plant physiology, soil ecology, fluxes of minerals and nutrients, and overall ecosystem function. The simplest system utilizes growth chambers and specialized root chambers with organic-less media to study the physiology of plant-mycorrhizal associations. A second system incorporates natural soil in open-top chambers to study soil bacterial and fungal population response to stress. The most complex CEF incorporates reconstructed soil profiles in a "constructed" ecosystem, enabling close examination of the soil foodweb. Our results show that closed ecosystem research is important for understanding mechanisms of response to ecosystem stresses. In addition, responses observed at one level of biological complexity may not allow prediction of response at a different level of biological complexity. In closed life support systems, incorporating soil foodwebs will require less artificial manipulation to maintain system stability and sustainability.     

Particulated growth media for optimal liquid and gaseous fluxes to plant roots in microgravity.

An important and yet relatively under researched area of plant growth in microgravity, deals with the rooting environment of plants. A comprehensive approach for selecting the physical characteristics of root growth media which optimizes the dynamic availability of water and dissolved nutrients, and gases to plant roots was developed and tested. Physically-based and parametric models describing the relationship between content and fluxes of liquids and gases were used to cast a multi-objective optimization problem. This methodology reveals that a medium's ability to supply liquid and gas fluxes optimally is dependent upon physiological target values, system operation limits and root module design which dictate the medium's range of soil water characteristic and particle size distribution. Optimized media parameters designate a particle size distribution from which a particulated growth media was constructed and matched to the optimized media parameters. This methodology should improve the selection of optimal media properties for plant growth in microgravity as well as other porous media applications.     

Use of lunar regolith as a substrate for plant growth.

Regenerative Life Support Systems (RLSS) will be required to regenerate air, water, and wastes, and to produce food for human consumption during long-duration missions to the Moon and Mars. It may be possible to supplement some of the materials needed for a lunar RLSS from resources on the Moon. Natural materials at the lunar surface may be used for a variety of lunar RLSS needs, including (i) soils or solid-support substrates for plant growth, (ii) sources for extraction of essential, plant-growth nutrients, (iii) substrates for microbial populations in the degradation of wastes, (iv) sources of O2 and H2, which may be used to manufacture water, (v) feed stock materials for the synthesis of useful minerals (e.g., molecular sieves), and (vi) shielding materials surrounding the outpost structure to protect humans, plants, and microorganisms from harmful radiation. Use of indigenous lunar regolith as a terrestrial-like soil for plant growth could offer a solid support substrate, buffering capacity, nutrient source/storage/retention capabilities, and should be relatively easy to maintain. The lunar regolith could, with a suitable microbial population, play a role in waste renovation; much like terrestrial waste application directly on soils. Issues associated with potentially toxic elements, pH, nutrient availability, air and fluid movement parameters, and cation exchange capacity of lunar regolith need to be addressed before lunar materials can be used effectively as soils for plant growth.     

A capillary-driven root module for plant growth in microgravity.

A new capillary-driven root module design for growing plants in microgravity was developed which requires minimal external control. Unlike existing systems, the water supply to the capillary-driven system is passive and relies on root uptake and media properties to develop driving gradients which operate a suction-induced flow control valve. A collapsible reservoir supplies water to the porous membrane which functions to maintain hydraulic continuity. Sheet and tubular membranes consisting of nylon, polyester and sintered porous stainless steel were tested. While finer pore sized membranes allow greater range of operation, they also reduce liquid flux thereby constraining system efficiency. Membrane selection should consider both the maximum anticipated liquid uptake rate and maximum operating matric head (suction) of the system. Matching growth media water retention characteristics to the porous membrane characteristics is essential for supplying adequate liquid flux and gas exchange. A minimum of 10% air-filled porosity (AFP) was necessary for adequate aeration. The capillary-driven module maintained hydraulic continuity and proper gas exchange rates for more than 80 days in a plant growth experiment.     

Development and research program for a soil-based bioregenerative agriculture system to feed a four person crew at a Mars base.

For humans to survive during long-term missions on the Martian surface, bioregenerative life support systems including food production will decrease requirements for launch of Earth supplies, and increase mission safety. It is proposed that the development of "modular biospheres"--closed system units that can be air-locked together and which contain soil-based bioregenerative agriculture, horticulture, with a wetland wastewater treatment system is an approach for Mars habitation scenarios. Based on previous work done in long-term life support at Biosphere 2 and other closed ecological systems, this consortium proposes a research and development program called Mars On Earth(TM) which will simulate a life support system designed for a four person crew. The structure will consist of 6 x 110 square meter modular agricultural units designed to produce a nutritionally adequate diet for 4 people, recycling all air, water and waste, while utilizing a soil created by the organic enrichment and modification of Mars simulant soils. Further research needs are discussed, such as determining optimal light levels for growth of the necessary range of crops, energy trade-offs for agriculture (e.g. light intensity vs. required area), capabilities of Martian soils and their need for enrichment and elimination of oxides, strategies for use of human waste products, and maintaining atmospheric balance between people, plants and soils.     

Measurements of trace contaminants in closed-type plant cultivation chambers.

Trace contaminants generated in closed facilities can cause abnormal plant growth. We present measurement data of trace contaminants released from soils, plants, and construction materials. We mainly used two closed chambers, a Closed-type Plant and Mushroom Cultivation Chamber (PMCC) and Closed-type Plant Cultivation Equipment (CPCE). Although trace gas budgets from soils obtained in this experiment are only one example, the results indicate that the budgets of trace gases, as well as CO2 and O2, change greatly with the degree of soil maturation and are dependent on the kind of substances in the soil. Both in the PMCC and in the CPCE, trace gases such as dioctyl phthalate (DOP), dibutyl phthalate (DBP), toluene and xylene were detected. These gases seemed to be released from various materials used in the construction of these chambers. The degree of increase in these trace gas levels was dependent on the relationship between chamber capacity and plant quantity. Results of trace gas measurement in the PMCC, in which lettuce and shiitake mushroom were cultivated, showed that ethylene was released both from lettuce and from the mushroom culture bed. The release rates were about 90 ng bed-1 h-1 for the shiitake mushroom culture bed (volume is 1700 cm3) and 4.1 approximately 17.3 ng dm-2 h-1 (leaf area basis) for lettuce. Higher ethylene release rates per plant and per unit leaf area were observed in mature plants than in young plants.     

Plants + soil/wetland microbes: Food crop systems that also clean air and water

The limitations that will govern bioregenerative life support applications in space, especially volume and weight, make multi-purpose systems advantageous. This paper outlines two systems which utilize plants and associated microbial communities of root or growth medium to both produce food crops and clean air and water. Underlying these approaches are the large numbers and metabolic diversity of microbes associated with roots and found in either soil or other suitable growth media. Biogeochemical cycles have microbial links and the ability of microbes to metabolize virtually all trace gases, whether of technogenic or biogenic origin, has long been established. Wetland plants and the rootzone microbes of wetland soils/media also been extensively researched for their ability to purify wastewaters of a great number of potential water pollutants, from nutrients like N and P, to heavy metals and a range of complex industrial pollutants. There is a growing body of research on the ability of higher plants to purify air and water. Associated benefits of these approaches is that by utilizing natural ecological processes, the cleansing of air and water can be done with little or no energy inputs. Soil and rootzone microorganisms respond to changing pollutant types by an increase of the types of organisms with the capacity to use these compounds. Thus living systems have an adaptive capacity as long as the starting populations are sufficiently diverse. Tightly sealed environments, from office buildings to spacecraft, can have hundreds or even thousands of potential air pollutants, depending on the materials and equipment enclosed. Human waste products carry a plethora of microbes which are readily used in the process of converting its organic load to forms that can be utilized by green plants. Having endogenous means of responding to changing air and water quality conditions represents safety factors as these systems operate without the need for human intervention. We review this research and the ability of systems using these mechanisms to also produce food or other useful crops. Concerns about possible pathogens in soils and wastewater are discussed along with some methods to prevent contact, disease transmission and to pre-screen and decrease risks. The psychological benefits of having systems utilizing green plants are becoming more widely recognized. Some recent applications extending the benefits of plants and microbes to solve new environmental problems are presented. For space applications, we discuss the use of in situ space resources and ways of making these systems compact and light-weight.     

Plant mineral nutrition,gas exchange and photosynthesis in space: A review

Successful growth and development of higher plants in space rely on adequate availability and uptake of water and nutrients, and efficient energy distribution through photosynthesis and gas exchange. In the present review, literature has been reviewed to assemble the relevant knowledge within space plant research for future planetary missions. Focus has been on fractional gravity, space radiation, magnetic fields and ultimately a combined effect of these factors on gas exchange, photosynthesis and transport of water and solutes.     

Sweetpotato vine management for confined food production in a space life-support system

Sweetpotato (Ipomea batatas L.) ‘Whatley–Loretan’ was developed for space life support by researchers at Tuskegee University for its highly productive, nutritious storage roots. This promising candidate space life-support crop has a sprawling habit and aggressive growth rate in favorable environments that demands substantial growing area. Shoot pruning is not a viable option for vine control because removal of the main shoot apex drastically inhibits storage-root initiation and development, and chemical growth retardants typically are not cleared for use with food crops. As part of a large effort by the NASA Specialized Center of Research and Training in Advanced Life Support to reduce equivalent system mass (ESM) for food production in space, the dilemma of vine management for sweetpotato was addressed in effort to conserve growth area without compromising root yield. Root yields from unbranched vines trained spirally around wire frames configured either in the shapes of cones or cylinders were similar to those from vines trained horizontally along the bench, but occupying only a small fraction of the bench area. This finding indicates that sweetpotato is highly adaptable to a variety of vine-training architectures. Planting a second plant in the growth container and training the two vines in opposite directions around frames enhanced root yield and number, but had little effect on average length of each vine or bench area occupied. Once again, root yields were similar for both configurations of wire support frames. The 3–4-month crop-production cycles for sweetpotato in the greenhouse spanned all seasons of multiple years during the course of the study, and although electric lighting was used for photoperiod control and to supplement photosynthetic light during low-light seasons, there still were differences in total light available across seasons. Light variations and other environmental differences among experiments in the greenhouse had more effects on vine length than on root yield. Average vine length correlated positively with total hours of daylight received across seasons, and responses for one plant per container were higher above a threshold duration of solar exposure, suggesting that the vines of two plants per container compete for available light. In addition to the adaptability of sweetpotato to various vine-training architectures and across seasons in terms of maintaining root productivity, the open, interior volumes of the support frames tested in this study will provide future opportunity to enhance sweetpotato root yield in space by adding novel interior lighting, such as from intracanopy arrays of light-emitting diodes. This work was sponsored by NASA grant NAG 5 1286.     

Current concepts and future directions of CELSS.

Studies of bioregenerative life support systems for use in space indicate that they are scientifically feasible. Preliminary data suggest that they would provide cost- and weight-saving benefits for low Earth orbit, long duration space platforms. Concepts of such systems include the use of higher plants and/or micro-algae as sources of food, potable water and oxygen, and as sinks for carbon dioxide and metabolic wastes. Recycling of materials within the system will require processing of food organism and crew wastes using microbiological and/or physical chemical techniques. The dynamics of material flow within the system will require monitoring, control, stabilization and maintenance imposed by computers. Future phases of study will continue investigations of higher plant and algal physiology, environmental responses, and control; flight experiments for testing responses of organisms to weightlessness and increased radiation levels; and development of ground-based facilities for the study of recycling within a bioregenerative life support system.     

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.     

Potato growth in a porous tube water and nutrient delivery system.

Potato (Solanum tuberosum L.) cv. 'Norland', vegetative growth and tuber productivity grown in the porous water and nutrient delivery system (PTNDS) developed by the Wisconsin Center for Space Automation and Robotics were compared with the vegetative growth and tuber productivity of plants grown in a peat:vermiculite potting mixture (PT/VR). The plants were grown at 12, 16, and 24-h light periods, 18 degrees C constant temperature, 70% relative humidity, and 300 micromol m-2 s-1 photosynthetic photon flux. Canopy height of plants grown in the PT/VR system was taller than that of plants grown in the PTNDS system. Canopy height differences were greatest when the plants were grown under a 24-h photoperiod. Leaf and stem dry masses were similar for plants grown in the two systems under the 12-h photoperiod. Under the 24-h photoperiod, leaf and stem dry masses of plants grown in the PT/VR system were more than 3 times those of plants grown in the PTNDS system. Tuber dry masses were similar for plants grown in the two systems under the 12-h photoperiod. Under the 24 h-photoperiod, tuber dry weights of plants grown in the PT/VR system were more than twice those of plants grown in the PTNDS system. A slightly higher harvest index (ratio of tuber weight to leaf plus stem weight) was noted for the plants grown in the PTNDS than for the plants grown in the PT/VR system. Plants grown in the PTNDS system at the 24-h photoperiod matured earlier than plants grown at this photoperiod in the PT/VR system. Vegetative growth and tuber productivity of plants grown under the 16-h photoperiod generally were intermediate to those noted for plants grown under the 12 and 24-h photoperiods. These results indicate that potato plants grown in a PTNDS system may require less plant growing volume, mature in a shorter time, and likely produce more tubers per unit area compared with plants grown in the PT/VR system. These plant characteristics are a distinct advantage for a plant growing unit of a CELSS.     

Development of life support requirements for long-term space flight.

When humans move out into the solar system to stay for long durations, the most immediate challenge will be the provision of a life-supporting environment in locations that are naturally devoid of food, air, and water. Life support systems must provide these commodities in all phases of space flight--during intravehicular activity (IVA) and during extra-vehicle activity (EVA). Systems that support human life must provide: overall reliability in the space environment, allowing maintenance and component replacement in space; reduced resupply mass of consumables and spares; for planetary surfaces, the ability to utilize local resources for increased self sufficiency; and the minimized mass power and volume requirements necessary for all space flight systems. This paper will discuss the melding of these technical requirements in such a way as to meet the human needs of space flight.     

Calcium balance in pea root statocytes under both clinorotation and Ca2+ channel blockers' influence.

We have previously demonstrated that space flight and clinorotation conditions increase cytoplasmic Ca2+ level in pea root statocytes. A rise in [Ca2+]i may be a serious problem for plants in microgravity environment. It is hypothesized that involvement of Ca2+ channel blockers in the growth medium may rescue a plant from abundance of Ca2+ ions. Indeed, combination of clinorotation (2 rpm, 5 days) and any Ca2+ channel blocker (1 micromole D600 or nicardipine, 12 hr) causes decreasing the Ca2+ concentration in pea root statocytes in comparison with clinorotation alone. Redistribution of Ca(2+)-ATPase activities observed under clinorotation comes to normal after D600 application whereas following by nicardipine action the pattern of the cytochemical staining is intermediate between those in stationary control and under clinorotation. Our data support the hypothesis that Ca2+ channel blockers may act as protectors for plants against rise in [Ca2+]i. The role for Ca2+ channels in graviperception and in microgravity effects as well as ways for stabilization of Ca2+ balance in plant cells in space flights are discussed.     

Seed-to-seed growth of Arabidopsis thaliana on the International Space Station.

The assembly of the International Space Station (ISS) as a permanent experimental outpost has provided the opportunity for quality plant research in space. To take advantage of this orbital laboratory, engineers and scientists at the Wisconsin Center for Space Automation and Robotics (WCSAR), University of Wisconsin-Madison, developed a plant growth facility capable of supporting plant growth in the microgravity environment. Utilizing this Advanced Astroculture (ADVASC) plant growth facility, an experiment was conducted with the objective to grow Arabidopsis thaliana plants from seed-to-seed on the ISS. Dry Arabidopsis seeds were anchored in the root tray of the ADVASC growth chamber. These seeds were successfully germinated from May 10 until the end of June 2001. Arabidopsis plants grew and completed a full life cycle in microgravity. This experiment demonstrated that ADVASC is capable of providing environment conditions suitable for plant growth and development in microgravity. The normal progression through the life cycle, as well as the postflight morphometric analyses, demonstrate that Arabidopsis thaliana does not require the presence of gravity for growth and development.     

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.     

A conceptual configuration of the lunar base bioregenerative life support system including soil-like substrate for growing plants

The paper presents a conceptual configuration of the lunar base bioregenerative life support system (LBLSS), including soil-like substrate (SLS) for growing plants. SLS makes it possible to combine the processes of plant growth and the utilization of plant waste. Plants are to be grown on SLS on the basis of 20 kg of dry SLS mass or 100 kg of wet SLS mass per square meter. The substrate is to be delivered to the base ready-made as part of the plant growth subsystem. Food for the crew was provided by prestored stock 24% and by plant growing system 76%. Total dry weight of the food is 631 g per day (2800 kcal/day) for one crew member (CM). The list of candidate plants to be grown under lunar BLSS conditions included 14 species: wheat, rice, soybean, peanuts, sweet pepper, carrots, tomatoes, coriander, cole, lettuce, radish, squash, onion and garlic. From the prestored stock the crew consumed canned fish, iodinated salt, sugar, beef sauce and seafood sauce. Our calculations show that to provide one CM with plant food requires the area of 47.5 m². The balance of substance is achieved by the removal dehydrated urine 59 g, feces 31 g, food waste 50 g, SLS 134 g, and also waters 86 g from system and introduction food 236 g, liquid potassium soap 4 g and mineral salts 120 g into system daily. To reduce system setup time the first plants could be sowed and germinated to a certain age on the Earth.     

Touch and gravitropic set-point angle interact to modulate gravitropic growth in roots.

Plant roots must sense and respond to a variety of environmental stimuli as they grow through the soil. Touch and gravity represent two of the mechanical signals that roots must integrate to elicit the appropriate root growth patterns and root system architecture. Obstacles such as rocks will impede the general downwardly directed gravitropic growth of the root system and so these soil features must be sensed and this information processed for an appropriate alteration in gravitropic growth to allow the root to avoid the obstruction. We show that primary and lateral roots of Arabidopsis do appear to sense and respond to mechanical barriers placed in their path of growth in a qualitatively similar fashion. Both types of roots exhibited a differential growth response upon contacting the obstacle that directed the main axis of elongation parallel to the barrier. This growth habit was maintained until the obstacle was circumvented, at which point normal gravitropic growth was resumed. Thus, the gravitational set-point angle of the primary and lateral roots prior to encountering the barrier were 95 degrees and 136 degrees respectively and after growing off the end of the obstacle identical set-point angles were reinstated. However, whilst tracking across the barrier, quantitative differences in response were observed between these two classes of roots. The root tip of the primary root maintained an angle of 136 degrees to the horizontal as it traversed the barrier whereas the lateral roots adopted an angle of 154 degrees. Thus, this root tip angle appeared dependent on the gravitropic set-point angle of the root type with the difference in tracking angle quantitatively reflecting differences in initial set-point angle. Concave and convex barriers were also used to analyze the response of the root to tracking along a continuously varying surface. The roots maintained the a fairly fixed angle to gravity on the curved surface implying a constant resetting of this tip angle/tracking response as the curve of the surface changed. We propose that the interaction of touch and gravity sensing/response systems combine to strictly control the tropic growth of the root. Such signal integration is likely a critical part of growth control in the stimulus-rich environment of the soil.