Modeling gravity effects on water retention and gas transport characteristics in plant growth substrates

Growing plants to facilitate life in outer space, for example on the International Space Station (ISS) or at planned deep-space human outposts on the Moon or Mars, has received much attention with regard to NASA’s advanced life support system research. With the objective of in situ resource utilization to conserve energy and to limit transport costs, native materials mined on Moon or Mars are of primary interest for plant growth media in a future outpost, while terrestrial porous substrates with optimal growth media characteristics will be useful for onboard plant growth during space missions. Due to limited experimental opportunities and prohibitive costs, liquid and gas behavior in porous substrates under reduced gravity conditions has been less studied and hence remains poorly understood. Based on ground-based measurements, this study examined water retention, oxygen diffusivity and air permeability characteristics of six plant growth substrates for potential applications in space, including two terrestrial analogs for lunar and Martian soils and four particulate substrates widely used in reduced gravity experiments. To simulate reduced gravity water characteristics, the predictions for ground-based measurements (1 − g) were scaled to two reduced gravity conditions, Martian gravity (0.38 − g) and lunar gravity (0.16 − g), following the observations in previous reduced gravity studies. We described the observed gas diffusivity with a recently developed model combined with a new approach that estimates the gas percolation threshold based on the pore size distribution. The model successfully captured measured data for all investigated media and demonstrated the implications of the poorly-understood shift in gas percolation threshold with improved gas percolation in reduced gravity. Finally, using a substrate-structure parameter related to the gaseous phase, we adequately described the air permeability under reduced gravity conditions.     

Gravitational effects on plant growth hormone concentration.

Numerous studies, particularly those of H. Dolk in the 1930's, established by means of bio-assay, that more growth hormone diffused from the lower, than from the upper side of a gravity-stimulated plant shoot. Now, using an isotope dilution assay, with 4,5,6,7 tetradeutero indole-3-acetic acid as internal standard, and selected ion monitoring-gas chromatography-mass spectrometry as the method of determination, we have confirmed Dolk's finding and established that the asymmetrically distributed hormone is, in fact, indole-3-acetic acid (IAA). This is the first physico-chemical demonstration that there is more free IAA on the lower sides of a geo-stimulated plant shoot. We have also shown that free IAA occurs primarily in the conductive vascular tissues of the shoot, whereas IAA esters predominate in the growing cortical cells. Now, using an especially sensitive gas chromatographic isotope dilution assay we have found that the hormone asymmetry also occurs in the non-vascular tissue. Currently, efforts are directed to developing isotope dilution assays, with picogram sensitivity, to determine how this asymmetry of IAA distribution is attained so as to better understand how the plant perceives the geo-stimulus.     

The effects of HGMFs on the plant gravisensing system.

High Gradient Magnetic Fields (HGMFs) offer new opportunities for studying the gravitropic system of plants. However, it is necessary to analyze the influence that HGMF can have on cellular processes and structures that may not be related to amyloplasts displacement. This paper considers possible HGMF effects on plants, which may accompany HGMF stimulation of amyloplasts and contribute to the mechanisms of the HGMF-induced curvature.     

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.     

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.     

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.     

The Breadboard Project: a functioning CELSS plant growth system.

The primary objective of the Breadboard project for the next 3-4 years is to develop, integrate and operate a Controlled Ecological Life Support System (CELSS) at a one person scale. The focus of this project over the past two years has been the development of the plant growth facility, the first module of the CELSS. The other major modules, food preparation, biomass processing, and resource recovery, have been researched at the laboratory scale during the past two years and facilities are currently under construction to scale-up these modules to an operational state. This paper will outline the design requirements for the Biomass Production Chamber (BPC), the plant growth facility for the project, and the control and monitoring subsystems which operate the chamber and will present results from both engineering and biological tests of the facility. Three production evaluations of wheat, conducted in the BPC during the past year, will be described and the data generated from these tests discussed. Future plans for the BPC will be presented along with future goals for the project as the other modules become active.     

In vitro plant cell growth in microgravity and on clinostat.

For the study of gravity's role in the processes of plant cell differentiation in-vitro, a model "seed-seedling-callus" has been used. Experiments were carried out on board the orbital stations Salyut-7 and Mir as well as on clinostat. They lasted from 18 to 72 days. It was determined that the exclusion of a one-sided action of gravity vector by means of clinostat and spaceflight conditions does not impede the formation and growth of callus tissue; however, at cell and subcellular levels structural and functional changes do take place. No significant changes were observed either on clinostat or in space concerning the accumulation of fresh biomass, while the percentage of dry material in space is lower than in control. Both in microgravity (MG) and in control, even after 72 days of growth, cells with a normally developed ultrastructure are present. In space, however, callus tissue more often contains cells in which the cross-section area of a cell, a nuclei and of mitochondria are smaller and the vacuole area--bigger than in controls. In microgravity a considerable decrease in the number of starch-containing cells and a reduction in the mean area of starch grains in amyloplasts is observed. In space the amount of soluble proteins in callus tissue is 1.5 times greater than in control. However, no differences were observed in fractions when separated by the SDS-PAGE method. In microgravity the changes in cell wall material components was noted. In the space-formed callus changes in the concentration of ions K, Na, Mg, Ca and P were observed. However, the direction of these changes depends on the age of callus. Discussed are the possible reasons for modification of morphological and metabolic parameters of callus cells when grown under changed gravity conditions.     

Instrumentation for plant health and growth.

Comprehensive spectroscopic monitoring of plant health and growth in bioregenerative life support system environments is possible using a variety of spectrometric technologies. Absorption spectrometry and atomic emission spectrometry in combination allow for direct, on-line, reagentless monitoring of plant nutrients from nitrate and potassium to micronutrients such as copper and zinc. Fluorometric spectrometry is ideal for the on-line detection, identification and quantification of bacteria and fungi. Liquid Atomic Emission Spectrometry (LAES) is a new form of spectrometry that allows for direct measurement of atomic emission spectra in liquids. An electric arc is generated by a pair of electrodes in the liquid to provide the energy necessary to break molecular bonds and reduce the substance to atomic form. With a fiber probe attached to the electrodes, spectral light can be transmitted to a photodiode array spectrometer for light dispersion and analysis. Ultraviolet (UV) absorption spectrometry is a long-established technology, but applications typically have required specific reagents to produce an analyte-specific absorption. Nitrate and iron nutrients have native UV absorption spectra that have been used to accurately determine nutrient concentrations at the +/- 5% level. Fluorescence detection and characterization of microbes is based upon the native fluorescent signatures of most microbiological species. Spectral and time-resolved fluorometers operating with remote fiber-optic probes will be used for on-line microbial monitoring in plant nutrient streams.     

Achieving and documenting closure in plant growth facilities.

As NASA proceeds with its effort to develop a Controlled Ecological Life Support System (CELSS) that will provide life support to crews during long duration space missions, it must address the question of facility and system closure. Here we discuss the concept of closure as it pertains to CELSS and describe engineering specifications, construction problems and monitoring procedures used in the development and operation of a closed plant growth facility for the CELSS program. A plant growth facility is one of several modules required for a CELSS. A prototype of this module at Kennedy Space Center is the large (7m tall x 3.5m diameter) Biomass Production Chamber (BPC), the central facility of the CELSS Breadboard Project. The BPC is atmospherically sealed to a leak rate of approximately 5% of its total volume per 24 hours. This paper will discuss the requirements for atmospheric closure in this facility, present CO2 and trace gas data from initial tests of the BPC with and without plants, and describe how the chamber was sealed atmospherically. Implications that research conducted in this type of facility will have for the CELSS program are discussed.     

Initial approach to comparative studies on the evolutionary potentials of space radiation effects in a plant system.

The role of cosmic ionizing radiation, including heavy ions (HZE-particles) in the induction of mutations at the molecule-, chromosome-, genome- and cell-level is discussed on the basis of different DNA organization in a pro- and eukaryotically compartmented plant system (Arabidopsis thaliana (L.) Heynh.). Data recently obtained on the biological effects of ionizing radiation make it timely to discuss comparatively the evolutionary potentials of space radiation effects in the pro- and eukaryotic genomes (plasmon, plastidom, chondriom, and nucleom) during long duration exposure on space flights.     

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.     

Transpiration during life cycle in controlled wheat growth.

We use a previously-developed model of wheat growth, which was designed for convenient incorporation into system-level models of advanced space life support systems. We apply the model to data from an experiment that grew wheat under controlled conditions and measured fresh biomass and cumulated transpiration as a function of time. We examine the adequacy of modeling the transpiration as proportional to the inedible biomass and an age factor, which varies during the life cycle. Results indicate that during the main phase of vegetative growth in the first half of the life cycle, the rate of transpiration per unit mass of inedible biomass is more than double the rate during the phase of grain development and maturation during latter half of the life cycle.     

Evaluating and optimizing horticultural regimes in space plant growth facilities.

In designing innovative space plant growth facilities (SPGF) for long duration space flight, various limitations must be addressed including onboard resources: volume, energy consumption, heat transfer and crew labor expenditure. The required accuracy in evaluating on board resources by using the equivalent mass methodology and applying it to the design of such facilities is not precise. This is due to the uncertainty of the structure and not completely understanding the properties of all associated hardware, including the technology in these systems. We present a simple criteria of optimization for horticultural regimes in SPGF: Qmax = max [M x (EBI)2/(V x E x T], where M is the crop harvest in terms of total dry biomass in the plant growth system; EBI is the edible biomass index (harvest index), V is volume occupied by the crop; E is the crop light energy supply during growth; T is the crop growth duration. The criterion reflects directly on the consumption of onboard resources for crop production.     

Interaction of a mixed yeast culture in an "autotroph-heterotroph" system with a closed atmosphere cycle and spatially separated components.

The study considers an experimental model of the "autotroph-heterotroph" system with a closed atmosphere cycle, in which the heterotrophic link is a mixed yeast population. The autotrophic link is represented by the algae Chlorella vulgaris and the heterotrophic link by the yeasts Candida utilis and Candida guilliermondii. The controls are populations of Chlorella and the same yeasts isolated from the atmosphere. It has been shown that the outcome of competition in the heterotrophic link depends on the strategy of the yeast population towards the substrate and oxygen. The C. utilis population quickly utilizes the substrate as it is an r-strategist and is less sensitive to oxygen deficiency. The C. guilliermondii population consumes low concentrations of the substrate because it is a K-strategist, but it is more sensitive to oxygen deficiency. That is why, in the "autotroph-heterotroph" system with a closed gas cycle, after a considerable amount of the substrate has been consumed, the C. guilliermondii population becomes more competitive that the C. utilis population. In the culture of yeasts, isolated from the atmosphere, the C. utilis population finds itself in more favorable conditions due to oxygen deficiency. The system with a complex heterotrophic component survive longer than a system whose heterotrophic component is represented by only one yeast species. This is explained for by the positive metabolite interaction of yeasts and a more complete utilization of the substrate by a mixed culture of yeasts featuring different strategies towards the substrate.     

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.     

Modeling gas exchange in a closed plant growth chamber.

Fluid transport models for fluxes of water vapor and CO2 have been developed for one crop of wheat and three crops of soybean grown in a closed plant growth chamber. Correspondence among these fluxes is discussed. Maximum fluxes of gases are provided for engineering design requirements of fluid recycling equipment in growth chambers. Furthermore, to investigate the feasibility of generalized crop models, dimensionless representations of water vapor fluxes are presented. The feasibility of such generalized models and the need for additional data are discussed.     

Optical sensors for monitoring and control of plant growth systems.

Optical chemical sensors have been developed for monitoring several parameters relevant to plant growth systems. These sensors utilize porous polymer and porous glass as the sensing element, and optical fiber input/output lines connected to a custom optoelectronic interface. Present in the sensing element are immobilized colorimetric indicators, which react with the analyte to be sensed. This reaction results in a change in the optical properties of the sensor. These sensors are particularly suited to in-situ monitoring of nutrient solution parameters and atmospheric trace contaminants in life support and plant growth systems. Sensors for monitoring pH, ammonia, and ethylene will be discussed.     

Solid matrix and liquid culture procedures for growth of potatoes.

This report discusses the advantages and limitations of several different procedures for growth of potatoes for CELSS. Solution culture, in which roots and stolons are submerged, and aeroponic culture were not found useful for potatoes because stolons did not produce tubers unless a severe stress was applied to the plants. In detailed comparison studies, three selected culture systems were compared, nutrient film technique (NFT), NFT with shallow media, and pot culture with deep media. For the NFT and NFT plus shallow media, plants were grown in 0.3 m2 trays and for the deep medium culture, in 20 liter pots. A 1 cm depth of arcillite, a baked montmorillonite clay, was used as shallow media (NFT-arc). Peat-vermiculite mixture was used to fill the pots for the deep media. Nutrient solution, modified half-strength Hoagland's, was recirculated among the tray culture plants with pH automatically controlled at 5.5, and conductivity maintained at approximately 1100 microS cm-1 by adding stock nutrients or renewing the solution. A separate nutrient solution was used to water the pot plants four times daily to excess and the excess was discarded. Plants of Norland cv. were utilized and transplanted from sterile-propagated stem cutting plantlets. The plants were grown for 66 days under 12 h photoperiod in a first study and grown for 54 days under 24 h photoperiod in a second study. Under both photoperiods, total plant growth was greater in NFT-arc than in either NFT or pot culture. Under 12 h photoperiod, tuber dry weight was 30% higher with NFT-arc, but 50% lower with NFT, than with pot culture. Under 24 h photoperiod, however, tuber dry weight in both NFT and NFT-arc was only 20% of that in pot culture. The NFT and NFT-arc produced a greater shoot growth and larger number of small tubers than pot culture, especially with 24 h photoperiod. It is concluded that there are serious limitations to the use of NFT alone for growth of potatoes in a CELSS system. These limitations can be minimized by using a modified NFT with a shallow layer of media, such as arcillite, yet additional work is needed to ensure high tuber production with this system under long photoperiods.