Composition and physical properties of starch in microgravity-grown plants.

The effect of spaceflight on starch development in soybean (Glycine max L., BRIC-03) and potato (Solanum tuberosum, Astroculture-05) was compared with ground controls by biophysical and biochemical measurements. Starch grains from plants from both flights were on average 20-50% smaller in diameter than ground controls. The ratio delta X/delta rho (delta X --difference of magnetic susceptibilities, delta rho--difference of densities between starch and water) of starch grains was ca. 15% and 4% higher for space-grown soybean cotyledons and potato tubers, respectively, than in corresponding ground controls. Since the densities of particles were similar for all samples (1.36 to 1.38 g/cm3), the observed difference in delta X/delta rho was due to different magnetic susceptibilities and indicates modified composition of starch grains. In starch preparations from soybean cotyledons (BRIC-03) subjected to controlled enzymatic degradation with alpha-amylase for 24 hours, 77 +/- 6% of the starch from the flight cotyledons was degraded compared to 58 +/- 12% in ground controls. The amylose content in starch was also higher in space-grown tissues. The good correlation between the amylose content and delta X/delta rho suggests, that the magnetic susceptibility of starch grains is related to their amylose content. Since the seedlings from the BRIC-03 experiment showed elevated post-flight ethylene levels, material from another flight experiment (GENEX) which had normal levels of ethylene was examined and showed no difference to ground controls in size distribution, density, delta X/delta rho and amylose content. Therefore the role of ethylene appears to be more important for changes in starch metabolism than microgravity.     

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.     

Proximate nutritional composition of CELSS crops grown at different CO2 partial pressures.

Two CELSS candidate crops, soybean (Glycine max) and potato (Solanum tuberosum), were grown hydroponically in controlled environments maintained at carbon dioxide (CO2) partial pressures ranging from 0.05 to 1.00 kPa (500 to 10,000 ppm at 101 kPa atmospheric pressure). Plants were harvested at maturity (90 days for soybean and 105 days for potato) and all tissues analyzed for proximate nutritional composition (i.e. protein, fat, carbohydrate, crude fiber, and ash content). Soybean seed ash and crude fiber were higher and carbohydrate was lower than values reported for field-grown seed. Potato tubers showed little difference from field-grown tubers. With the exception of increased crude fiber of soybean seed with increased CO2, no trends were apparent with regard to CO2 effects on proximate composition of soybean seed and potato tubers. Crude fiber of soybean stems and leaves increased with increased CO2, as did soybean leaf protein (total nitrogen). Potato leaf and stem (combined) protein levels also increased with increased CO2, while leaf and stem carbohydrates decreased. Values for leaf and stem protein and ash were higher than values generally reported for field-grown plants for both species. Results suggest that CO2 partial pressure should have little influence on proximate composition of potato tubers or soybean seed, but that high ash and protein levels might be expected from leaves and stems of crops grown in controlled environments of a CELSS.     

Selection and hydroponic growth of potato cultivars for bioregenerative life support systems

As part of the ESA-funded MELiSSA program, Ghent University and the Université catholique de Louvain investigated the suitability, growth and development of four potato cultivars in hydroponic culture under controlled conditions with the aim to incorporate such cultivation system in an Environmental Control and Life Support System (ECLSS). Potato plants can fulfill three major functions in an ECLSS in space missions: (a) fixation of CO₂ and production of O₂, (b) production of tubers for human nutrition and (c) production of clean water after condensation of the water vapor released from the plants by transpiration. Four cultivars (Annabelle, Bintje, Desiree and Innovator) were selected and grown hydroponically in nutrient film technique (NFT) gullies in a growth chamber under controlled conditions. The plant growth parameters, tuber harvest parameters and results of tuber nutritional analysis of the four cultivars were compared. The four potato cultivars grew well and all produced tubers. The growth period lasted 127 days for all cultivars except for Desiree which needed 145 days. Annabelle (1.45 kg/m²) and Bintje (1.355 kg/m²) were the best performing of the four cultivars. They also produced two times more tubers than Desiree and Innovator. Innovator produced the biggest tubers (20.95 g/tuber) and Desiree the smallest (7.67 g/tuber). The size of Annabelle and Bintje potatoes were intermediate. Bintje plants produced the highest total biomass in term of DW. The highest non-edible biomass was produced by Desiree, which showed both the highest shoot and root DW. The manual length and width measurements were also used to predict the total tuber mass. The energy values of the tubers remained in the range of the 2010 USDA and Souci-Fachmann-Kraut food composition databases. The amount of Ca determined was slightly reduced compared to the USDA value, but close to the Souci-Fachmann-Kraut value. The concentration of Cu, Zn and P were high compared to both databases.     

Structure of cress root statocytes in microgravity (Bion-10 mission).

Experiments on primary roots of Lepidium sativum L. have been performed on board the Bion-10 satellite. The experimental set-up was extremely miniaturized and completely automatic. The results demonstrate the effectiveness of the instrumentation. The spatial orientation, growth, root cap differentiation and statocyte structure of roots grown under microgravity (MG) have been compared with control roots grown on the ground (GC) and in a 1G-reference centrifuge in space (RC). Root length and cap shape did not differ between MG and control samples. Under MG, the mean distance of the statoliths from the distal cell wall of the statocytes increased significantly, the mean distance of the mitochondria decreased and the nucleus did not change its position in comparison to both controls. The number and the shape of the amyloplasts (statoliths) were not influenced by the space flight factors, but their size as well as their relative area in the cell decreased. The number of starch grains per statolith as well as their size and shape changed under MG. In MG and RC samples the number of lipid bodies in the statocytes was higher and the relative area larger than in GC samples. The relative area occupied by vacuoles was greater in RC statocytes than in GC and MG statocytes. These results partly confirm and, in addition, extend the data from earlier experiments in space.     

Use of biologically reclaimed minerals for continuous hydroponic potato production in a CELSS.

Plant-derived nutrients were successfully recycled in a Controlled Ecological Life Support System (CELSS) using biological methods. The majority of the essential nutrients were recovered by microbiologically treating the plant biomass in an aerobic bioreactor. Liquid effluent containing the nutrients was then returned to the biomass production component via a recirculating hydroponic system. Potato (Solanum tuberosum L.) cv. Norland plants were grown on those nutrients in either a batch production mode (same age plants on a nutrient solution) or a staggered production mode (4 different ages of plants on a nutrient solution). The study continued over a period of 418 days, within NASA Breadboard Project's Biomass Production Chamber at the Kennedy Space Center. During this period, four consecutive batch cycles (104-day harvests) and 13 consecutive staggered cycles (26-day harvests) were completed using reclaimed minerals and compared to plants grown with standard nutrient solutions. All nutrient solutions were continually recirculated during the entire 418 day study. In general, tuber yields with reclaimed minerals were within 10% of control solutions. Contaminants, such as sodium and recalcitrant organics tended to increase over time in solutions containing reclaimed minerals, however tuber composition was comparable to tubers grown in the control solutions.     

Approaches in the determination of plant nutrient uptake and distribution in space flight conditions.

The effective growth and development of vascular plants rely on the adequate availability of water and nutrients. Inefficiency in either the initial absorption, transportation, or distribution of these elements are factors which impinge on plant structure and metabolic integrity. The potential effect of space flight and microgravity conditions on the efficiency of these processes is unclear. Limitations in the available quantity of space-grown plant material and the sensitivity of routine analytical techniques have made an evaluation of these processes impractical. However, the recent introduction of new plant cultivating methodologies supporting the application of radionuclide elements and subsequent autoradiography techniques provides a highly sensitive investigative approach amenable to space flight studies. Experiments involving the use of gel based 'nutrient packs' and the radionuclides calcium-45 and iron-59 were conducted on the Shuttle mission STS-94. Uptake rates of the radionuclides between ground and flight plant material appeared comparable.     

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.     

Excess nutrients in hydroponic solutions alter nutrient content of rice, wheat, and potato.

Environment has significant effects on the nutrient content of field-grown crop plants. Little is known, however, about compositional changes caused by controlled environments in which plants receive only artificial radiation and soilless, hydroponic culture. This knowledge is essential for developing a safe, nutritious diet in a Controlled Ecological Life-Support System (CELSS). Three crops that are candidates for inclusion in a CELSS (rice, wheat, and white potato) were grown both in the field and in controlled environments where the hydroponic nutrient solution, photosynthetic photon flux (PPF), and CO2 level were manipulated to achieve rapid growth rates. Plants were harvested at maturity, separated into discrete parts, and dried prior to analysis. Plant materials were analyzed for proximate composition (protein, fat, ash, and carbohydrate), total nitrogen (N), nitrate, minerals, and amino-acid composition. The effect of environment on nutrient content varied by crop and plant part. Total N and nonprotein N (NPN) contents of plant biomass generally increased under controlled-environment conditions compared to field conditions, especially for leafy plant parts and roots. Nitrate levels were increased in hydroponically-grown vegetative tissues, but nitrate was excluded from grains and tubers. Mineral content changes in plant tissue included increased phosphorus and decreased levels of certain micronutrient elements under controlled-environment conditions. These findings suggest that cultivar selection, genetic manipulation, and environmental control could be important to obtain highly nutritious biomass in a CELSS.     

Spaceflight experiments with Arabidopsis starch-deficient mutants support a statolith-based model for graviperception.

In order to help resolve some of the controversy associated with ground-based research that has supported the starch-statolith theory of gravity perception in plants, we performed spaceflight experiments with Arabidopsis in Biorack during the January 1997 and May 1997 missions of the Space Shuttle. Seedlings of wild-type (WT) Arabidopsis, two reduced-starch strains, and a starchless mutant were grown in microgravity and then were given either a 30, 60, or 90 minute gravity stimulus on a centrifuge. By the 90 min 1-g stimulus, the WT exhibited the greatest magnitude of curvature and the starchless mutant exhibited the smallest curvature while the two reduced starch mutants had an intermediate magnitude of curvature. In addition, space-grown plants had two structural features that distinguished them from the controls: a greater number of root hairs and an anomalous hypocotyl hook structure. However, the morphological changes observed in the flight seedlings are likely to be due to the effects of ethylene present in the spacecraft. (Additional ground-based studies demonstrated that this level of ethylene did not significantly affect gravitropism nor did it affect the relative gravitropic sensitivity among the four strains.) Nevertheless, this experiment on gravitropism was performed the "right way" in that brief gravitational stimuli were provided, and the seedlings were allowed to express the response without further gravity stimuli. Our spaceflight results support previous ground-based studies of these and other mutants since increasing amounts of starch correlated positively with increasing sensitivity to gravity.     

Somatic embryos of daylily in space

Poor growth and nuclear abnormalities observable in some space-grown plants have been hypothesized as due to a combination of factors such as degree of development, the specific way the plants are grown and the way they experience multiple stresses, some of which are space-specific. Data from a 132-day experiment on ‘Mir’ using embryogenic cell cultures of daylily (Hemerocallis) allow seemingly contradictory evidence from earlier Shuttle missions to be harmonized: a) the more developed an embryo the less likely it is to suffer catastrophic cell stress during growth, whereas the less developed it is, the greater its vulnerability; (b) the extent to which the stress becomes manifest is also dependent on the extent of pre-existing stresses imposed by suboptimal growing conditions; (c) an appropriate, albeit undesirable, ‘stress match’ with other non-equilibrium determinants, much like a ‘tug of war’, can result in genomic variations in space. It is not understood what is/are the feature(s) of the space environment that cause the various cell division perturbations but they have not yet been mimicked on earth. The stress symptoms were found only in space materials and, as predicted, they were most frequently encountered in smaller, less-developed materials grown under non-optimized conditions. It is concluded that, while any substantial deviation from ‘optimum’ can be a ‘stress’, spaceflight subjects vulnerable materials to cell division or DNA-repair stress(es) that appear distinctive, but remain elusive so far. Fastidiously-controlled growing environments must be devised to resolve the matter of direct versus indirect effects of space. On a practical level, it is predicted that adapting plant biotechnologies to space conditions will not be a casual matter.     

THE SPACE EXPERIMENT OF PROTEIN CRYSTALLIZATION ABOARD THE CHINESE SPACECRAFT SZ-3

Using a new flight hardware, a Chinese mission of space protein crystallization has been performed on the Chinese spacecraft SZ-3 for seven days from March 25 to Apr. 1, 2002. The rate of yielding crystals in the 60 samples is 75%, a little higher than the ground control experiment. Preliminary analysis of the experimental results have shown that among the 16 proteins involved in the mission, about 4 kinds of better diffraction-quality protein crystals were produced in space. At least one kind of protein crystals, i.e. crystals of cytochrome b5 mutant could diffract X ray beyond the highest resolution reported so far. In addition, some rules derived from our numerical studies of the liquid/liquid diffusion protein crystallization were proved by the crystallization of lysozyme as model protein in this space experiment,which also clearly showed the advantages and disadvantages of the gelled protein solution used in microgravity growth of protein crystals. In order to exploit this mission, more diffraction work with the grown crystals and detailed analysis of data to be obtained will be done in the next few months.     

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.]     

Cytochemical localization of calcium in soybean root cap cells in microgravity.

The antimonate precipitation technique was used to evaluate the effects of microgravity and ethylene on the cellular and subcellular distribution of free calcium ions in soybean root apices. Soybean (Glycine max L. [Merr.]) dry seeds were launched, activated by hydration, and germinated in the presence of KMnO4 (to remove ethylene) and in its absence onboard the space shuttle Columbia during the STS-87 mission. Primary root apices of 6-day old seedlings were fixed for electron microscopy after landing. Ultrastructural studies indicated that antimonate precipitation appeared as individual electron-dense particles which were more or less round in shape and varied in diameter from 10 nm (minimum size beginning from which the particles were well identified) to 90 nm. It was revealed that analyzed root cap cells varied in both the precipitate particle sizes and the amount particles per unit of the cellular area. In both flight and ground control treatments, antimonate precipitation level increases from apical meristem cells to peripheral (secretory) cells of root apices. In root cap statocytes, subcellular localization of precipitate particles was revealed in the cytoplasm, nucleus and small vacuoles. The quantitative analysis showed a reduction of precipitate density in the cytoplasm and the nucleus, and an increase in precipitate density in the vacuoles from statocytes of both spaceflight treatments in comparison with ground controls.     

Modelling of cometary nuclei: Planetary missions preparation

The planning of planetary missions requires the knowledge as much as possible accurate of the target. This knowledge, which cannot be obtained only by ground based observations, can be supported by theoretical modeling. This is particularly true in the case of Rosetta, a cornerstone ESA mission that will be launched in January 2003, and of its target, the comet 46P/Wirtanen. In this paper we show how, using a nucleus thermal evolution model, it is possible to foresee the temperature of the nucleus surface and the activity level along the orbit and define the range of possible values for these properties. Activity level close to the aphelion depends on the presence of CO. Surface temperatures depend on the physical properties attributed to the dust: temperatures reached by a crust mainly composed by silicatic grains differ from the temperatures reached by a crust mainly composed by organic grains.     

Comparison of aerobically-treated and untreated crop residue as a source of recycled nutrients in a recirculating hydroponic system.

This study compared the growth of potato plants on nutrients recycled from inedible potato biomass. Plants were grown for 105 days in recirculating, thin-film hydroponic systems containing four separate nutrient solution treatments: (1) modified half-strength Hoagland's (control), 2) liquid effluent from a bioreactor containing inedible potato biomass, 3) filtered (0.2 micrometer) effluent, and 4) the water soluble fraction of inedible potato biomass (leachate). Approximately 50% of the total nutrient requirement in treatments 2-4 were provided (recycled) from the potato biomass. Leachate had an inhibitory effect on leaf conductance, photosynthetic rate, and growth (50% reduction in plant height and 60% reduction in tuber yield). Plants grown on bioreactor effluent (filtered or unfiltered) were similar to the control plants. These results indicated that rapidly degraded, water soluble organic material contained in the inedible biomass, i.e., material in leachate, brought about phytotoxicity in the hydroponic culture of potato. Recalcitrant, water soluble organic material accumulated in all nutrient recycling treatments (650% increase after 105 days), but no increase in rhizosphere microbial numbers was observed.     

Skeletogenesis in sea urchin larvae under modified gravity conditions.

From many points of view, skeletogenesis in sea urchins has been well described. Based on this scientific background and considering practical aspects of sea urchin development (i.e. availability of material, size of larvae, etc.), we wanted to know whether orderly skeletogenesis requires the presence of gravity. The objective has been approached by three experiments successfully performed under genuine microgravity conditions (in the STS-65 IML-2 mission of 1994; in the Photon-10 IBIS mission of 1995 and in the STS-76 S/MM-03 mission of 1996). Larvae of the sea urchin Sphaerechinus granularis were allowed to develop in microgravity conditions for several days from blastula stage onwards (onset of skeletogenesis). At the end of the missions, the recovered skeletal structures were studied with respect to their mineral composition, architecture and size. Live larvae were also recovered for post-flight culture. The results obtained clearly show that the process of mineralisation is independent of gravity: that is, the skeletogenic cells differentiate correctly in microgravity. However, abnormal skeleton architectures were encountered, particularly in the IML-2 mission, indicating that the process of positioning of the skeletogenic cells may be affected, directly or indirectly, by environmental factors, including gravity. Larvae exposed to microgravity from blastula to prism/early pluteus stage for about 2 weeks (IBIS mission), developed on the ground over the next 2 months into normal metamorphosing individuals.     

Integrating biological treatment of crop residue into a hydroponic sweetpotato culture.

Residual biomass from hydroponic culture of sweetpotato [Ipomoea batatas (L.) Lam.] was degraded using natural bacterial soil isolates. Sweetpotato was grown for 120 days in hydroponic culture with a nutrient solution comprised of a ratio of 80% modified half Hoagland solution to 20% filtered effluent from an aerobic starch hydrolysis bioreactor. The phytotoxicity of the effluent was assayed with Waldmann's Green' lettuce (Lactuca sativa L.) and the ratio selected after a 60-day bioassay using sweetpotato plants propagated vegetatively from cuttings. Controlled environment chamber experiments were conducted to investigate the impact of filtrate from biological treatment of crop residue on growth and storage root production with plants grown in a modified half Hoagland solution. Incorporation of bioreactor effluent, reduced storage root yield of 'Georgia Jet' sweetpotato but the decrease was not statistically significant when compared with yield for plants cultured in a modified half Hoagland solution without filtrate. However, yield of 'TU-82-155' sweetpotato was significantly reduced when grown in a modified half Hoagland solution into which filtered effluent had been incorporated. Total biomass was significantly reduced for both sweetpotato cultivars when grown in bioreactor effluent. The leaf area and dry matter accumulation were significantly (P < 0.05) reduced for both cultivars when grown in solution culture containing 20% filtered effluent.     

Characterizing parameters of Jatropha curcas cell cultures for microgravity studies

Jatropha (Jatropha curcas) is a tropical perennial species identified as a potential biofuel crop. The oil is of excellent quality and it has been successfully tested as biodiesel and in jet fuel mixes. However, studies on breeding and genetic improvement of jatropha are limited. Space offers a unique environment for experiments aiming at the assessment of mutations and differential gene expression of crops and in vitro cultures of plants are convenient for studies of genetic variation as affected by microgravity. However, before microgravity studies can be successfully performed, pre-flight experiments are necessary to characterize plant material and validate flight hardware environmental conditions. Such preliminary studies set the ground for subsequent spaceflight experiments. The objectives of this study were to compare the in vitro growth of cultures from three explant sources (cotyledon, leaf, and stem sections) of three jatropha accessions (Brazil, India, and Tanzania) outside and inside the petriGAP, a modified group activation pack (GAP) flight hardware to fit petri dishes. In vitro jatropha cell cultures were established in petri dishes containing a modified MS medium and maintained in a plant growth chamber at 25 ± 2 °C in the dark. Parameters evaluated were surface area of the explant tissue (A), fresh weight (FW), and dry weight (DW) for a period of 12 weeks. Growth was observed for cultures from all accessions at week 12, including subsequent plantlet regeneration. For all accessions differences in A, FW and DW were observed for inside vs. outside the PetriGAPs. Growth parameters were affected by accession (genotype), explant type, and environment. The type of explant influenced the type of cell growth and subsequent plantlet regeneration capacity. However, overall cell growth showed no abnormalities. The present study demonstrated that jatropha in vitro cell cultures are suitable for growth inside PetriGAPs for a period of 12 weeks. The parameters evaluated in this study provide the basic ground work and pre-flight assessment needed to justify a model for microgravity studies with jatropha in vitro cell cultures. Future studies should focus on results of experiments performed with jatropha in vitro cultures in microgravity.     

Effect of CO2 levels on nutrient content of lettuce and radish.

Atmospheric carbon-dioxide enrichment is known to affect the yield of lettuce and radish grown in controlled environments, but little is known about CO2 enrichment effects on the chemical composition of lettuce and radish. These crops are useful model systems for a Controlled Ecological Life-Support System (CELSS), largely because of their relatively short production cycles. Lettuce (Lactuca sativa L.) cultivar 'Waldmann's Green' and radish (Raphanus sativus L.) cultivar 'Giant White Globe' were grown both in the field and in controlled environments, where hydroponic nutrient solution, light, and temperature were regulated, and where CO2 levels were controlled at 400, 1000, 5000, or 10,000 ppm. Plants were harvested at maturity, dried, and analyzed for proximate composition (protein, fat, ash, and carbohydrate), total nitrogen (N), nitrate N, free sugars, starch, total dietary fiber, and minerals. Total N, protein N, nonprotein N (NPN), and nitrate N generally increased for radish roots and lettuce leaves when grown under growth chamber conditions compared to field conditions. The nitrate-N level of lettuce leaves, as a percentage of total NPN, decreased with increasing levels of CO2 enrichment. The ash content of radish roots and of radish and lettuce leaves decreased with increasing levels of CO2 enrichment. The levels of certain minerals differed between field- and chamber-grown materials, including changes in the calcium (Ca) and phosphorus (P) contents of radish and lettuce leaves, resulting in reduced Ca/P ratio for chamber-grown materials. The free-sugar contents were similar between the field and chamber-grown lettuce leaves, but total dietary fiber content was much higher in the field-grown plant material. The starch content of growth-chamber lettuce increased with CO2 level.