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At least 19 records

Gravity independence of seed-to-seed cycling in Brassica rapa

Growth of higher plants in the microgravity environment of orbital platforms has been problematic. Plants typically developed more slowly in space and often failed at the reproductive phase. Short-duration experiments on the Space Shuttle showed that early stages in the reproductive process could occur normally in microgravity, so we sought a long-duration opportunity to test gravity's role throughout the complete life cycle. During a 122-d opportunity on the Mir space station, full life cycles were completed in microgravity with Brassica rapa L. in a series of three experiments in the Svet greenhouse. Plant material was preserved in space by chemical fixation, freezing, and drying, and then compared to material preserved in the same way during a high-fidelity ground control. At sampling times 13 d after planting, plants on Mir were the same size and had the same number of flower buds as ground control plants. Following hand-pollination of the flowers by the astronaut, siliques formed. In microgravity, siliques ripened basipetally and contained smaller seeds with less than 20% of the cotyledon cells found in the seeds harvested from the ground control. Cytochemical localization of storage reserves in the mature embryos showed that starch was retained in the spaceflight material, whereas protein and lipid were the primary storage reserves in the ground control seeds. While these successful seed-to-seed cycles show that gravity is not absolutely required for any step in the plant life cycle, seed quality in Brassica is compromised by development in microgravity.

Flight Experiment

Effect of cultural conditions on the seed-to-seed growth of Arabidopsis and Cardamine - A study of growth rates and reproductive development as affected by test tube seals

The effects of test tube seals on the growth, flowering, and seed pod formation of Arabidopsis thaliana (L.) Heynh., mouse ear cress, and Cardamine oligosperma Nutt, bitter cress, are studied in order to assess the conditions used in weightlessness experiments. Among other results, it is found that the growth (height) and flowering (date of bud appearance) were suppressed in mouse ear cress in tubes sealed with Saran. Seed pod formation which occurred by day 45 in open-to-air controls, was still lacking in the sealed plants even up to day 124. The growth and flowering of bitter cress were also suppressed by the Saran seal, although up to day 55 the Saran-sealed plants were taller. It is suggested that atmospheric composition was the cause of the suppression of growth, flowering, and seed pod development in these plants, since the mouse ear cress renewed their growth and then set seed pods after the Saran seal was ruptured.

Hoshizaki, T.

Seed-to-seed growth of superdwarf wheat and arabidopsis using red light-emitting diodes (LED's): A report on baseline tests conducted for NASA's proposed Plant Research Unit (PRU)

To determine the influence of narrow-spectrum red light-emitting diodes (LED's) on plant growth and seed production, wheat (Triticum aestivum L.cv Superdwarf) and Arabidopsis (Arabidopsis thaliana (L.) Heynh, race Columbia) plants were grown under red LED's (peak emission 660 nm) and compared to plants grown under daylight fluorescent (white) light and red LED's supplemented with either 1 percent or 10 percent blue fluorescent (BF) light. Wheat growth under red LED's alone appeared normal, whereas Arabidopsis under red LED's alone developed curled leaf margins and a spiraling growth pattern. Both wheat and Arabidopsis under red LED's alone or red LED's + 1 percent BF light had significantly lower seed yield than plants grown under white light. However, the addition of 10 percent BF light to red LED's partially alleviated the adverse effect of red LED's on yield. Irrespective of the light treatment, viable seeds were produced by wheat(75-92 percent germination rate) and Arabidopsis (85-100 percent germination rate). These results indicate that wheat, and to a lesser extent Arabidopsis, can be successfully grown under red LED's alone, but supplemental blue light is required with red LED's to sufficiently match the growth characteristics and seed yield associated with plants grown under white light.

Goins, G. D.

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. c2003 COSPAR. Published by Elsevier Ltd. All rights reserved.

ISS Project

Greenhouse (III): Gas-Exchange and Seed-to-Seed Experiments on the Russian Space Station MIR and Earth-grown, Ethylene-Treated Wheat Plants

The Mir Space Station provided an outstanding opportunity to study long-term plant responses when exposed to a microgravity environment. Furthermore, if plants can be grown to maturity in a microgravity environment, they might be used in future bioregenerative life-support systems (BLSS). The primary objective of the Greenhouse experiment onboard Mir was to grow Super Dwarf and Apogee wheat through complete life cycles in microgravity; i.e., from seed-to-seed-to-seed. Additional objectives were to study chemical, biochemical, and structural changes in plant tissues as well as photosynthesis, respiration, and transpiration (evaporation of water from plants). Another major objective was to evaluate the suitability clothe facilities on Mir for advanced research with plants. The Greenhouse experiment was conducted in the Russian/Bulgarian plant growth chamber, the Svet, to which the United States added instrumentation systems to monitor changes in CO2 and water vapor caused by the plants (with four infrared gas analyzers monitoring air entering and leaving two small plastic chambers). In addition, the US instrumentation also monitored O2; air, leaf (IR), cabin pressure; photon flux; and substrate temperature and substrate moisture (16 probes in the root module). Facility modifications were first performed during the summer of 1995 during Mir 19, which began after STS-72 left Mir. Plant development was monitored by daily observations and some photographs.

Campbell, William F.

[Growth of wheat from seed-to-seed in space flight]

In our earlier space experiment with super dwarf wheat we found the spikes developed in space to be barren. The cause of the full crop sterility was sensitivity of this wheat species to the ethylene concentration of 0.3-0.8 mg/m3 during the experiment. The follow-up ground experiments were made to identify species of dwarf wheat that could be raised in space greenhouse Svet and are distinguished by partial tolerance of their reproductive organs to elevated ethylene in air. The choice fell on the USU-Apogee cultivar specially developed for planting in growth chambers as an integral part of various bioregenerative life support systems, including the space ones. An experiment with wheat Apogee was performed in greenhouse Svet on board MIR. The period of the full crop vegetation cycle was not significantly altered under the spaceflight conditions. The experiment yielded 508 seeds from 12 plants, i.e. by 38% less than in laboratory experiments and by 69% more as compared with results of growing crops in ethylene-contaminated atmosphere (1 mg/m3). Mass of the space seeds was low if compared with the laboratory crops. This was the first time when the feasibility of gathering seeds from wheat that had passed the whole vegetation cycle in space flight was demonstrated. The experiment will give mightly impetus to the advancement of research on space biological LSS and gravitational biology.

long duration

In vitro seed to seed growth on clinostats

The effect of a long term micro-gravity environment on the life cycle of plants is unknown. Whether higher plants have evolved to a stage where removal or reduction of gravity is detrimental to plant life cycle and thus fatal to the plant species, is an unanswered question in space plants which were successfully grown through the various stages of their life cycle. Attempts to grow plants as a continuous integral process from seed to seed through one generation were successful until recently. Culture of plants through multiple generations was not accomplished in space nor in ground based studies. The effect of long term simulated weightlessness by growing consecutive generations of plants continuously on clinostats using the cruciferous plants, Arabidopsis thaliana (L.) Heyn. and Cardamine oligosperma Nutt. is being investigated.

Hoshizaki, T.

X-Ray Characterization of Structural Defects in Seeded and Self-Seeded ZnSe Crystal Grown by PVT in Horizontal and Vertical Configurations

As part of a pre-flight ground based investigation of crystal growth of II-VI compound semiconductors, a number of ZnSe boules have been grown by physical vapor transport (PVT) at Marshall Space Flight Center. Boules were grown in both horizontal and vertical configurations and seeded and self-seeded growth techniques were employed. As-grown and/or cleaved boules were examined by a combination of synchrotron white beam x-ray topography (SWBXT) and high resolution triple axis diffraction (HRTXD) to characterized the structural defects and correlate them with the growth conditions. Horizontal grown boules tend to grow away from the ampoule wall (contactless growth) and generally exhibit large (110) facets parallel to the gravity vector. Vertical grown boules grew to the full diameter of the ampoule and exhibited no faceting. X-ray topography combined with back reflection x-ray diffraction revealed the presence of lamellar twins (180 deg type about the [111] axis) in horizontal grown boules while vertically grown boules contain a few large grains, some of which are twinned. X-ray topographs and reciprocal space maps recorded from the boules show the better crystal quality of horizontal grown boules. The relationship between crystal quality and gravity vector is investigated. Further, an attempt is made to extend the Hurle theory of twin nucleation in Czochralski grown crystals to explain the twinning mechanisms in horizontal grown boules.

Raghothamachar, B.

Automated seed manipulation and planting

The Mechanical Division fabricated three seed separators utilizing pressure gradients to move and separate wheat seeds. These separators are called minnow buckets and use air, water, or a combination of both to generate the pressure gradient. Electrostatic fields were employed in the seed separator constructed by the Electrical Division. This separator operates by forcing a temporary electric dipole on the wheat seeds and using charged electrodes to attract and move the seeds. Seed delivery to the hydroponic growth tray is accomplished by the seed cassette. The cassette is compatible with all the seed separators, and it consists of a plastic tube threaded with millipore filter paper. During planting operations, the seeds are placed in an empty cassette. The loaded cassette is then placed in the growth tray and nutrient solution provided. The solution wets the filter paper and capillary action draws the nutrients up to feed the seeds. These seeding systems were tested and showed encouraging results. Seeds were effectively separated and the cassette can support the growth of wheat plants. Problems remaining to be investigated include improving the success of delivering the seeds to the cassette and providing adequate spacing between seeds for the electric separator.

Garcia, Ray

Continued results of the seeds in space experiment

Two million seeds of 120 different varieties representing 106 species, 97 genera, and 55 plant families were flown aboard the Long Duration Exposure Facility (LDEF). The seed were housed on the Space Exposed Experiment Developed for Students (SEEDS) tray in the sealed canister number 6 and in two small vented canisters. The tray was in the F-2 position. The seed were germinated and the germination rates and the development of the resulting plants were compared to the performance of the control seed that stayed in Park Seed's seed storage facility. The initial results were presented in a paper at the First LDEF Post-Retrieval Symposium. There was a better survival rate of the seed in the sealed canister in space than in the storage facility at Park Seed. At least some of the seed in each of the vented canisters survived the exposure to vacuum for almost six years. The number of observed apparent mutations was very low. In the initial testing, the small seeded crops were not grown to maturity to check for mutations and obtain second generation seed. These small seeded crops have now been grown for evaluation and second generation seed collected.

Alston, Jim A.

Germination, growth rates, and electron microscope analysis of tomato seeds flown on the LDEF

The tomato seeds were flown in orbit aboard the Long Duration Exposure Facility (LDEF) for nearly six years. During this time, the tomato seeds received an abundant exposure to cosmic radiation and solar wind. Upon the return of the LDEF to earth, the seeds were distributed throughout the United States and 30 foreign countries for analysis. The purpose of the experiment was to determine the long term effect of cosmic rays on living tissue. Our university analysis included germination and growth rates as well as Scanning Electron Microscopy and X-ray analysis of the control as well as Space-exposed tomato seeds. In analyzing the seeds under the Electron Microscope, usual observations were performed on the nutritional and epidermis layer of the seed. These layers appeared to be more porous in the Space-exposed seeds than on the Earth-based control seeds. This unusual characteristic may explain the increases in the space seeds growth pattern. (Several test results show that the Space-exposed seeds germinate sooner than the Earth-Based seeds. Also, the Space-exposed seeds grew at a faster rate). The porous nutritional region may allow the seeds to receive necessary nutrients and liquids more readily, thus enabling the plant to grow at a faster rate. Roots, leaves and stems were cut into small sections and mounted. After sputter coating the specimens with Argon/Gold Palladium Plasma, they were ready to be viewed under the Electron Microscope. Many micrographs were taken. The X-ray analysis displayed possible identifications of calcium, potassium, chlorine, copper, aluminum, silicon, phosphate, carbon, and sometimes sulfur and iron. The highest concentrations were shown in potassium and calcium. The Space-exposed specimens displayed a high concentration of copper and calcium in the two specimens. There was a significantly high concentration of copper in the Earth-based specimens, whereas there was no copper in the Space-exposed specimens.

Hammond, Ernest C., Jr.

Dynamics of storage reserve deposition during Brassica rapa L. pollen and seed development in microgravity

Pollen and seeds share a developmental sequence characterized by intense metabolic activity during reserve deposition before drying to a cryptobiotic form. Neither pollen nor seed development has been well studied in the absence of gravity, despite the importance of these structures in supporting future long-duration manned habitation away from Earth. Using immature seeds (3-15 d postpollination) of Brassica rapa L. cv. Astroplants produced on the STS-87 flight of the space shuttle Columbia, we compared the progress of storage reserve deposition in cotyledon cells during early stages of seed development. Brassica pollen development was studied in flowers produced on plants grown entirely in microgravity on the Mir space station and fixed while on orbit. Cytochemical localization of storage reserves showed differences in starch accumulation between spaceflight and ground control plants in interior layers of the developing seed coat as early as 9 d after pollination. At this age, the embryo is in the cotyledon elongation stage, and there are numerous starch grains in the cotyledon cells in both flight and ground control seeds. In the spaceflight seeds, starch was retained after this stage, while starch grains decreased in size in the ground control seeds. Large and well-developed protein bodies were observed in cotyledon cells of ground control seeds at 15 d postpollination, but their development was delayed in the seeds produced during spaceflight. Like the developing cotyledonary tissues, cells of the anther wall and filaments from the spaceflight plants contained numerous large starch grains, while these were rarely seen in the ground controls. The tapetum remained swollen and persisted to a later developmental stage in the spaceflight plants than in the ground controls, even though most pollen grains appeared normal. These developmental markers indicate that Brassica seeds and pollen produced in microgravity were physiologically younger than those produced in 1 g. We hypothesize that microgravity limits mixing of the gaseous microenvironments inside the closed tissues and that the resulting gas composition surrounding the seeds and pollen retards their development.

short duration

Space Exposed Experiment Developed for Students (SEEDS) (P0004-2)

SEEDS, a cooperative endeavor of NASA Headquarters, the NASA Langley Research Center, and the George W. Park Seed Company, resulted in the distribution of approximately 132,000 SEEDS kits to 3.3 million students. Kits contained Rutger's tomato seeds that had flown on the Long Duration Exposure Facility (LDEF), as well as seeds that had been stored in a climate controlled warehouse for the same time period. Preliminary data indicates the germination rate for space exposed seeds was 73.8 percent while Earth based seeds germinated at a rate of 70.3 percent. Tests conducted within the first six months after retrieval indicated space exposed seeds germinated in an average of 8.0 days, while Earth based seeds' average germination time was 8.3 days. Some mutations (assumed to be radiation induced) include plants that added a leaf instead of the usual flower at the end of the flower frond. Also, fruit produced from a flower with a variegated calyx bore seeds producing albino plants, while fruit from a flower with a green calyx from the same plant bore seeds producing green plants.

Grigsby, Doris K.

Cytochemical localization of reserves during seed development in Arabidopsis thaliana under spaceflight conditions

Successful development of seeds under spaceflight conditions has been an elusive goal of numerous long-duration experiments with plants on orbital spacecraft. Because carbohydrate metabolism undergoes changes when plants are grown in microgravity, developing seed storage reserves might be detrimentally affected during spaceflight. Seed development in Arabidopsis thaliana plants that flowered during 11 d in space on shuttle mission STS-68 has been investigated in this study. Plants were grown to the rosette stage (13 d) on a nutrient agar medium on the ground and loaded into the Plant Growth Unit flight hardware 18 h prior to lift-off. Plants were retrieved 3 h after landing and siliques were immediately removed from plants. Young seeds were fixed and processed for microscopic observation. Seeds in both the ground control and flight plants are similar in their morphology and size. The oldest seeds from these plants contain completely developed embryos and seed coats. These embryos developed radicle, hypocotyl, meristematic apical tissue, and differentiated cotyledons. Protoderm, procambium, and primary ground tissue had differentiated. Reserves such as starch and protein were deposited in the embryos during tissue differentiation. The aleurone layer contains a large quantity of storage protein and starch grains. A seed coat developed from integuments of the ovule with gradual change in cell composition and cell material deposition. Carbohydrates were deposited in outer integument cells especially in the outside cell walls. Starch grains decreased in number per cell in the integument during seed coat development. All these characteristics during seed development represent normal features in the ground control plants and show that the spaceflight environment does not prevent normal development of seeds in Arabidopsis.

STS-68 Shuttle Project

Impact of Space Radiation on Plant Seeds: from Arabidopsis thaliana to Crops

One of the major concerns for long-term exploration missions beyond the Earth’s magnetosphere is radiation risk, primarily from solar particle events (SPE) and galactic cosmic rays (GCR). With the goal of manned Mars exploration, the production of fresh food during long-duration space missions provides critical nutritional supplementation and may also benefit astronauts’ behavioral health. However, the effects of space radiation on plants and plant propagules have not been sufficiently investigated and characterized. This is the final report of our project to summarize the findings of the effect of space radiation exposure on plant seeds from morphometrics, nutritional values, to molecular machenisms. In this study, we evaluated the effect of simulated GCR (using dry seeds) or SPE (using hydrated seeds) on seeds of Arabidopsis, Mizuna mustard, ‘Outredgeous’ red romaine lettuce, and ‘Red Robin’ dwarf tomato. Seeds were exposed to various doses of simulated space radiation scenarios, either acutely or at a low dose rate (LDR), using the NASA Space Radiation Laboratory (NSRL) facility at Brookhaven National Lab (BNL). Exposure to simulated GCR or SPE at the levels tested had no significant impact on the germination rate in Arabidopsis and crop seeds; however, GCR reduced the viability of lettuce and tomato seeds. Overall, the morphological changes of the seedlings cultured from irradiated seeds were dose- and ion quality- dependent, with heavier ions causing more severe damage. These changes ranged from cotyledon deformation, shortened root length, smaller seedling size, and other signs of stress, depending on the seed types. Both 40 and 80 cGy (LDR) exposures of GCR or SPE significantly affected tomato early seedling development, delayed tomato fruiting, and reduced the total yield of tomato. Altered nutritional values were also found in edible biomass, especially for the GCR 80 cGy (LDR) groups. Underlying mechanisms were furtherly evaluated using transcriptomic analysis. For both GCR and SPE, 40 cGy showed some effects, but to a much lesser extent compared with 80 cGy, which can be considered as the “maximum permissible exposure” for the seed types we evaluated in this study. The impact of space radiation on seeds potentially affects the ability of plants to adapt to other environmental stresses (e.g. microgravity, water stress, and hardware constraints) as well as susceptibility to plant diseases, which need to be furtherly investigated. This research is funded by NASA’s Human Research Program.

J T Richards

Impact of Space Radiation on Plant Seeds: from Arabidopsis thaliana to Crops

One of the major concerns for long-term exploration missions beyond the Earth’s magnetosphere is radiation risk, primarily from solar particle events (SPE) and galactic cosmic rays (GCR). With the goal of manned Mars exploration, the production of fresh food during long-duration space missions provides critical nutritional supplementation and may also benefit astronauts’ behavioral health. However, the effects of space radiation on plants and plant propagules have not been sufficiently investigated and characterized. This is the final report of our project to summarize the findings of the effect of space radiation exposure on plant seeds from morphometrics, nutritiaonal values, to molecular machenisms. In this study, we evaluated the effect of simulated GCR (using dry seeds) or SPE (using hydrated seeds) on seeds of Arabidopsis, Mizuna mustard, ‘Outredgeous’ red romaine lettuce, and ‘Red Robin’ dwarf tomato. Seeds were exposed to various doses of simulated space radiation scenarios, either acutely or at a low dose rate (LDR), using the NASA Space Radiation Laboratory (NSRL) facility at Brookhaven National Lab (BNL). Exposure to simulated GCR or SPE at the levels tested had no significant impact on the germination rate in Arabidopsis and crop seeds; however, GCR reduced the viability of lettuce and tomato seeds. Overall, the morphological changes of the seedlings cultured from irradiated seeds were dose- and ion quality- dependent, with heavier ions causing more severe damage. These changes ranged from cotyledon deformation, shortened root length, smaller seedling size, and other signs of stress, depending on the seed types. Both 40 and 80 cGy (LDR) exposures of GCR or SPE significantly affected tomato early seedling development, delayed tomato fruiting, and reduced the total yield of tomato. Altered nutritional values were also found in edible biomass, especially for the GCR 80 cGy (LDR) groups. Underlying mechanisms were furtherly evaluated using transcriptomic analysis. For both GCR and SPE, 40 cGy showed some effects, but to a much lesser extent compared with 80 cGy, which can be considered as the “maximum permissible exposure” for the seed types we evaluated in this study. The impact of space radiation on seeds potentially affects the ability of plants to adapt to other environmental stresses (e.g. microgravity, water stress, and hardware constraints) as well as susceptibility to plant diseases, which need to be furtherly investigated. This research is funded by NASA’s Human Research Program.

J.T. Richards

Seed surface sanitization and persistence of E.coli through different tissues of ‘Red Robin’ Tomato (Solanum lycopersicum cv. Red Robin)

Seed surface sanitization via chemical processes removes/reduces microbes from the external surfaces of the seed and thereby could have an impact on the plants’ health or productivity. To determine the impact of seed surface sanitization on the plants’ microbiome, sanitized and unsanitized seeds from ‘Red Robin’ Tomato (Solanumlycopersicum cv. Red Robin) were exposed to Escherichia coli (E. coli) and grown in a controlled environment growth chamber simulating environmental conditions aboard the International Space Station (ISS). Plants were harvested at four intervals, days 11, 33,42 and 76 post-germination. Changes in the microbial communities of leaf, stem, root, and fruit because of E. coli exposure and the persistence of E. coli itself were investigated using aerobic plate count (APC), qPCR and 16S rRNA sequencing. It was determined that E. coli persisted for longer periods of time in plants from sanitized versus unsanitized seeds and was identified in root tissue more frequently than in leaf or stem tissue. E. coli was not detected in fruits raised from either sanitized or unsanitized seeds. The 16S rRNA sequencing showed dynamic changes in the abundance of members of the phylum Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes in all tissue types studied. We observed minimal or no changes in the alpha diversity of leaf stem and fruit tissue with time, or between sanitized and unsanitized seeds. Roots showed significant differences in alpha diversity with time and seed sanitization status. Beta-diversity showed that time had more of an influence on all samples versus the E. coli treatment. Members of phyla Proteobacteria and Bacteroidetes were found to be differentially abundant across leaf, stem and root tissue. Our results indicated that the seed surface sanitization, although a requirement for sending seeds to space, might influence the developing microbiome. This research was funded by NASA’s Space Life and Physical Sciences Research and Applications.

Anirudha R Dixit