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Musgrave, M. E.

Publications and source records attributed to Musgrave, M. E..

26 records · Page 2

Plant reproduction in spaceflight environments

Because plant reproduction is a complex developmental process there are many possible sites of perturbation by the unusual environments of orbital spacecraft. Previous long-duration experiments on Soviet platforms shared features of slowed development through the vegetative stage of plant growth and aborted reproductive function. Our goal has been to understand how special features of the spaceflight environment impact physiological function and reproductive development. In a series of short-duration experiments in the Shuttle mid-deck we studied early reproductive development in Arabidopsis thaliana. Pollen and ovule development aborted at an early stage in the first experiment on STS-54 which utilized closed plant growth chambers. Post-flight analysis suggested that the plants may have been carbon dioxide limited. Subsequent experiments utilized carbon dioxide enrichment (on STS-51) and cabin air flow-through with an air exchange system (on STS-68). Both modifications allowed pollen and ovule development to occur normally on orbit, and full reproductive development up to the stage of an immature seed occurred on STS-68. However, analysis of plant roots from these experiments demonstrated a limitation in rootzone aeration in the spaceflight material that was not mitigated by these procedures. In the future, additional resources (crew time, upgraded flight hardware, and special platforms) will invite more elaborate, long-duration experimentation. On the ISS, a variable speed centrifuge and upgraded plant habitats will permit detailed experiments on the role of gravity in shaping the plant micro-environment, and what influence this plays during reproduction.

Review, Tutorial↗

Spaceflight exposure effects on transcription, activity, and localization of alcohol dehydrogenase in the roots of Arabidopsis thaliana

Although considerable research and speculation have been directed toward understanding a plant's perception of gravity and the resulting gravitropic responses, little is known about the role of gravity-dependent physical processes in normal physiological function. These studies were conducted to determine whether the roots of plants exposed to spaceflight conditions may be experiencing hypoxia. Arabidopsis thaliana (L.) Heynh. plants were grown in agar medium during 6 or 11 d of spaceflight exposure on shuttle missions STS-54 (CHROMEX-03) and STS-68 (CHROMEX-05), respectively. The analysis included measurement of agar redox potential and root alcohol dehydrogenase (ADH) activity, localization, and expression. ADH activity increased by 89% as a result of spaceflight exposure for both CHROMEX-03 and -05 experiments, and ADH RNase protection assays revealed a 136% increase in ADH mRNA. The increase in ADH activity associated with the spaceflight roots was realized by a 28% decrease in oxygen availability in a ground-based study; however, no reduction in redox potential was observed in measurements of the spaceflight bulk agar. Spaceflight exposure appears to effect a hypoxic response in the roots of agar-grown plants that may be caused by changes in gravity-mediated fluid and/or gas behavior.

STS-68 Shuttle Project↗

Modification of reproductive development in Arabidopsis thaliana under spaceflight conditions

Reproductive development in Arabidopsis thaliana (L.) Heynh. cv. Columbia plants was investigated under spaceflight conditions on shuttle mission STS-51. Plants launched just prior to initiation of the reproductive phase developed flowers and siliques during the 10-d flight. Approximately 500 flowers were produced in total by the 12 plants in both the ground control and spaceflight material, and there was no significant difference in the number of flowers in each size class. The flower buds and siliques of the spaceflight plants were not morphologically different from the ground controls. Pollen viability tests immediately post-flight using fluorescein diacetate indicated that about 35% of the pollen was viable in the spaceflight material. Light-microscopy observations on this material showed that the female gametophytes also had developed normally to maturity. However, siliques from the spaceflight plants contained empty, shrunken ovules, and no evidence of pollen transfer to stigmatic papillae was found by light microscopy immediately post-flight or by scanning electron microscopy on fixed material. Short stamen length and indehiscent anthers were observed in the spaceflight material, and a film-like substance inside the anther that connected to the tapetum appeared to restrict the release of pollen from the anthers. These observations indicate that given appropriate growing conditions, early reproductive development in A. thaliana can occur normally under spaceflight conditions. On STS-51, reproductive development aborted due to obstacles in pollination or fertilization.

manned↗

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↗

Dynamics of vegetative cytoplasm during generative cell formation and pollen maturation in Arabidopsis thaliana

Ultrastructural changes of pollen cytoplasm during generative cell formation and pollen maturation in Arabidopsis thaliana were studied. The pollen cytoplasm develops a complicated ultrastructure and changes dramatically during these stages. Lipid droplets increase after generative cell formation and their organization and distribution change with the developmental stage. Starch grains in amyloplasts increase in number and size during generative and sperm cell formation and decrease at pollen maturity. The shape and membrane system of mitochondria change only slightly. Dictyosomes become very prominent, and numerous associated vesicles are observed during and after sperm cell formation. Endoplasmic reticulum appears extensively as stacks during sperm cell formation. Free and polyribosomes are abundant in the cytoplasm at all developmental stages although they appear denser at certain stages and in some areas. In mature pollen, all organelles are randomly distributed throughout the vegetative cytoplasm and numerous small particles appear. Organization and distribution of storage substances and appearance of these small particles during generative and sperm cell formation and pollen maturation are discussed.

Non-NASA Center↗

Pollen and ovule development in Arabidopsis thaliana under spaceflight conditions

The development of pollen and ovules in Arabidopsis thaliana on the space shuttle 'Endeavour' (STS-54) was investigated. Plants were grown on nutrient agar for 14 days prior to loading into closed plant growth chambers that received light and temperature control inside the Plant Growth Unit flight hardware on the shuttle middeck. After 6 days in spaceflight the plants were retrieved and immediately dissected and processed for light and electron microscope observation. Reproductive development aborted at an early stage. Pistils were collapsed and ovules inside were seen to he empty. No viable pollen was observed from STS-54 plants; young microspores were deformed and empty. At a late stage, the cytoplasm of the pollen contracted and became disorganized, but the pollen wall developed and the exine appeared normal. The tapetum in the flight flowers degenerated at early stages. Ovules from STS-54 flight plants stopped growing and the integuments and nucellus collapsed and degenerated. The megasporocytes appeared abnormal and rarely underwent meiosis. Apparently they enlarged, or occasionally produced a dyad or tetrad, to assume the form of a female gametophyte with the single nucleus located in an egglike cell that lacks a cell wall. Synergids, polar nuclei, and antipodals were not observed. The results demonstrate the types of lesions occurring in plant reproductive material under spaceflight conditions.

STS-54 Shuttle Project↗

A growth analysis of waterlogging damage in mung bean (Phaseolus aureus)

Mung beans (Phaseolus aureus Roxb.) were grown for 2 weeks in gravel-vermiculite soilless mix in a growth chamber and subjected to a 1-week waterlogging period followed by a 1-week recovery period. Sequential harvests were made to determine the time course of effects of waterlogging and subsequent recovery on growth parameters by techniques of growth analysis. Root dry matter was the first to be affected, along with an increase in leaf dry matter and specific leaf weight. After a 1-week waterlogging period, specific leaf weight had more than doubled in the stressed plants. Leaf area declined in relation to the control plants as did the ratio of root dry matter to shoot dry matter. During the recovery period there was an increase in the dry matter allocation to the roots relative to the shoot. Specific leaf weight fell to control levels although the rate of leaf area elaboration did not increase during this time, suggesting a redistribution of stored assimilates from the leaves. Net assimilation rate increased during the waterlogging period, probably due to a restriction in root metabolism and reduced translocation out of the leaf rather than to an increase in photosynthesis. Net assimilation rate of waterlogged plants was severely reduced compared with control plants during the recovery period. Both relative growth rate and leaf area duration declined during the waterlogging period and declined further subsequent to the waterlogging treatment. The results illustrate the interrelationships between root and shoot carbon budgets in mung bean during response to the stress of waterlogging.

Non-NASA Center↗

Growth and mitochondrial respiration of mungbeans (Phaseolus aureus Roxb.) germinated at low pressure

Mungbean (Phaseolus aureus Roxb.) seedlings were grown hypobarically to assess the effects of low pressure (21-24 kilopascals) on growth and mitochondrial respiration. Control seedlings grown at ambient pressure (101 kilopascals) were provided amounts of O2 equivalent to those provided experimental seedlings at reduced pressure to factor out responses to O2 concentration and to total pressure. Respiration was assayed using washed mitochondria, and was found to respond only to O2 concentration. Regardless of total pressure, seedlings grown at 2 millimoles O2 per liter had higher state 3 respiration rates and decreased percentages of alternative respiration compared to ambient (8.4 millimoles O2 per liter) controls. In contrast, seedling growth responded to total pressure but not to O2 concentration. Seedlings were significantly larger when grown under low pressure. While low O2 (2 millimoles O2 per liter) diminished growth at ambient pressure, growth at low pressure in the same oxygen concentration was enhanced. Respiratory development and growth of mungbean seedlings under low pressure is unimpaired whether oxygen or air is used as the chamber gas, and further, low pressure can improve growth under conditions of poor aeration.

Non-NASA Center↗