Engineering topics

Guikema, James A.

Publications and source records attributed to Guikema, James A..

14 records

NASA NTRS2004

Brassica rapa plants adapted to microgravity with reduced photosystem I and its photochemical activity

The photosynthetic apparatus contains several protein complexes, many of which are regulated by environmental conditions. In this study, the influences of microgravity on PSI and PSII in Brassica rapa plants grown aboard the space shuttle were examined. We found that Brassica plants grown in space had a normal level of growth relative to controls under similar conditions on Earth. Upon return to Earth, cotyledons were harvested and thylakoid membranes were isolated. Analysis of chlorophyll contents showed that the Chl a/b ratio (3.5) in flight cotyledons was much higher than a ratio of 2.42 in the ground controls. The flight samples also had a reduction of PSI complexes and a corresponding 30% decrease of PSI photochemical activity. Immunoblotting showed that the reaction centre polypeptides of PSI were more apparently decreased (e.g. by 24-33% for PsaA and PsaB, and 57% for PsaC) than the light-harvesting complexes. In comparison, the accumulation of PSII complex was less affected in microgravity, thus only a slight reduction in D1, D2 and LHCII was observed in protein blots. However, there was a 32% decrease of OEC1 in the flight samples, indicating a defective OEC subcomplex. In addition, an average 54% increase of the 54 kDa CF1-beta isoform was found in the flight samples, suggesting that space-grown plants suffered from certain stresses, consistent with implications of the increased Chl a/b ratio. Taken together, the results demonstrated that Brassica plants can adapt to spaceflight microgravity, but with significant alterations in chloroplast structures and photosynthetic complexes, and especially reduction of PSI and its activity.

Non-NASA Center↗

NASA NTRS1999

Starch Metabolism in Space-Grown Soybean Seedlings

The research conducted during this grant is described. There were three major areas of study: These were: (1) the interaction of gravity and sugar metabolism in soybean; (2) the effects of gravity on the photosynthetic activity of Brassica rapa; (3) investigation as to the effects of microgravity on the interaction of a fungal root pathogen with soybean.

Guikema, James A.↗

NASA NTRS1999

Starch Metabolism in Space-Grown Soybean Seedlings

The focus of this research was the study of sugar metabolism in soybean plants that had been in a clinorotation condition. The scope of activities was broadened greatly after the onset of the award. This broadening added two major research foci: 1) B-PAC: Photosynthetic activity of Brassica rapa; and 2) SOYPAT: The effects of microgravity on the interaction of a fungal root pathogen with soybean. Substantial investment and activity was also focused on the training of the astronaut team to conduct these experiments during orbital spaceflight.

Guikema, James A.↗

NASA NTRS1995

Clinorotation affects soybean seedling morphology

Although spaceflight does not appear to significantly affect seed germination, it can influence subsequent plant growth. On STS-3 and SL-2, decreased growth (measured as plant length, fresh weight, and dry weight) was noted for pine, oat, and mung bean. In the CHROMEX-01 and 02 experiments with Haplopappus and in the CHROMEX-03 experiment with Arabidopsis, enhanced root growth was noted in the space-grown plants. In the CHROMEX-04 experiments with wheat, both leaf fresh weight and leaf area were diminished in the space-grown plants but there was no difference in total plant height (CS Brown, HG Levine, and AD Krikorian, unpublished data). These data suggest that microgravity impacts growth by whole plant partitioning of the assimilates. The objective of the present study was to determine the influence of clinorotation on the growth and the morphology of soybean seedlings grown in the Biological Research In Canister (BRIC) flight hardware. This experiment provided baseline data for a spaceflight experiment (BRIC-3) flown on STS-63 (February 3-11, 1995).

Hilaire, Emmanuel↗

NASA NTRS1995

Effects of stress ethylene inhibitors on sweet clover (Melilotus Alba L.) seedling growth in microgravity

Previous experiments from our lab have shown that seeds of sweet clover, when germinated and grown within the Fluid Processing Apparatus (FPA) on a slow rotating clinostat produce significantly greater levels of the volatile stress hormone, ethylene, when compared to seeds treated the same but without clinorotation. In both conditions, carbon dioxide levels reached high levels and seedling growth was inhibited. However, clinorotation inhibited growth to a greater extent. To help determine to what extent microgravity influences stress ethylene production and to what extent ethylene inhibits seedling growth, we have extended the above experiments by growing sweet clover in the presence of aminooxyacetic acid (AOA) and silver nitrate (AgNO3), inhibitors of stress ethylene biosynthesis and action, respectively. Seeds of sweet clover were germinated and grown for five days in the FPA under two gravity conditions: under stationary conditions on Earth and in microgravity onboard the space shuttle, Discovery (STS-63), which launched Feb. 3, 1995. Upon recovery, gas samples were aspirated from the growth chambers and carbon dioxide and ethylene concentrations were measured using a gas chromatograph. Then the tissue was weighed, photographed and fixed, and is current undergoing further morphological and microscopic characterization.

Gallegos, Gregory L.↗

NASA NTRS1995

Effects of microgravity and clinorotation on stress ethylene production in two starchless mutants of Arabidopsis thaliana

Starch filled plastids termed amyloplasts, contained within columella cells of the root caps of higher plant roots, are believed to play a statolith-like role in the gravitropic response of roots. Plants having amyloplasts containing less starch exhibit a corresponding reduction in gravitropic response. We have observed enhanced ethylene production by sweet clover (Melilotus alba L.) seedlings grown in the altered gravity condition of a slow rotating clinostat, and have suggested that this is a stress response resulting from continuous gravistimulation rather than as a result of the simulation of a microgravity condition. If so, we expect that plants deficient in starch accumulation in amyloplasts may produce less stress ethylene when grown on a clinostat. Therefore, we have grown Arabidopsis thaliana in the small, closed environment of the Fluid Processing Apparatus (FPA). In this preliminary report we compare stationary plants with clinorotated and those grown in microgravity aboard Discovery during the STS-63 flight in February 1995. In addition to wildtype, two mutants deficient in starch biosynthesis, mutants TC7 and TL25, which are, respectively, deficient in the activity of amyloplast phosphoglucomutase and ADP-glucose pyrophosphorylase, were grown for three days before being fixed within the FPA. Gas samples were aspirated from the growth chambers and carbon dioxide and ethylene concentations were measured using a gas chromatograph. The fixed tissue is currently undergoing further morphologic and microscopic characterization.

Gallegos, Gregory L.↗

NASA NTRS1995

Effect of microgravity on stress ethylene and carbon dioxide production in sweet clover (Melilotus alba L.)

The study of higher plant growth and development in the microgravity (micro-g) environment continues to be a challenge. This is in part a result of the available flight qualified hardware with restrictive closed gas environments. This point is underscored by considering that gas exchange of seedlings grown in microgravity may be further limited owing to a thicker layer of water wicked onto the roots and to the absence of convective mixing. We hypothesized that seedlings grown under such conditions will experience greater hypoxia in microgravity than at Earth gravity, and thus produce greater stress ethylene. We compared flight and ground samples of sweet clover seedlings grown in the Fluid Processing Apparatus (FPA) during STS-57 and found them to contain extremely high levels of carbon dioxide (CO2) and stress ethylene. There were time dependent increases for both gases, and seedling growth was greatly inhibited. We repeated these experiments aboard STS-60 using modified chambers which increased, by fifty fold, the air available to the developing seedlings. Sweet clover seed germination and subsequent seedling growth to eight days within the FPA modified with a gas permeable membrane is not compromised by the microgravity environment.

Gallegos, Gregory L.↗

NASA NTRS1995

Plasmid acquisition in microgravity

In microgravity, bacteria often show an increased resistance to antibiotics. Bacteria can develop resistance to an antibiotic after transformation, the acquisition of DNA, usually in the form of a plasmid containing a gene for resistance to one or more antibiotics. In order to study the capacity of bacteria to become resistant to antibiotics in microgravity, we have modified the standard protocol for transformation of Escherichia coli for use in the NASA-flight-certified hardware package, The Fluid Processing Apparatus (FPA). Here we report on the ability of E. coli to remain competent for long periods of time at temperatures that are readily available on the Space Shuttle, and present some preliminary flight results.

Juergensmeyer, Margaret A.↗

NASA NTRS1995

The Fluid Processing Apparatus: from Flight Hardware to Electron Micrographs

Since the early years of space biology, a major drawback in spaceflight plant experiments has been the inability to fix specimens in microgravity, relying instead on fixation after return to Earth. As there, it is of a growing interest to look at the effect of microgravity on the structure and the developmental polarity of root graviperceptive cells, or columella cells, and so, it is important to use flight hardware which allows specimen fixation in space therefore avoiding the confounding effects of rapid readaptation to gravity after landing. As part of the Bioserve Space Technologies, a Center for the Commercial Development of Space (CCDS), we now have experiment flight opportunities through the Commercial Generic Bioprocessing Apparatus (CGBA) payload. In this study the Fluid Processing Apparatus (FPA) was used to grow seedlings for a limited period of time prior to fixation of the tissue in a microgravity environment. Upon return to Earth, the samples were processed for electron microscopy. This report describes the microscopic data obtained from the two space flights (STS-54 and STS-60). In both cases, the electron micrographs of the columella cells revealed well preserved cell structure, well defined microtubules, and the presence of calcium precipitates formed by a antimonate precipitation method.

Hilaire, Emmanuel↗

NASA NTRS1994

Production and action of cytokines in space

B6MP102 cells, a continuously cultured murine bone marrow macrophage cell line, were tested for secretion of tumor necrosis factor-alpha and Interleukin-1 during space flight. We found that B6MP102 cells secreted more tumor necrosis factor-alpha and interleukin-1 when stimulated in space with lipopolysaccharide than controls similarly stimulated on earth. This compared to increased secretion of interferon-beta and -gamma by lymphocytes that was measured on the same shuttle flights. Although space flight enhanced B6MP102 secretion of tumor necrosis factor-alpha, an experiment on a subsequent space flight (STS-50) found that cellular cytotoxicity, mediated by tumor necrosis factor-alpha, was inhibited.

Chapes, Stephen K.↗

NASA NTRS1994

Clover development during spaceflight: A model system

The development of legume root nodules was studied as a model system for the examination of gravitational effects on plant root development. In order to examine whether rhizobial association with clover roots can be achieved in microgravity, experiments were performed aboard the KC-135 parabolic aircraft and aboard the sounding rocket mission Consort 3. Binding of rhizobia to roots and the initial stages of root nodule development successfully occurred in microgravity. Seedling germination experiments were performed in the sliding block device, the Materials Dispersion Apparatus, aboard STS-37. When significant hydration of the seeds was achieved, normal rates of germination and seedling development were observed.

Guikema, James A.↗

NASA NTRS1994

Educational opportunities within the NASA specialized center of research and training in gravitational biology

The NASA Specialized Center of Research and Training (NSCORT) in Gravitational Biology was established at Kansas State University, supported through NASA's Life Science Division, Office of Space Science and Applications. Educational opportunities, associated with each of the research projects which form the nucleus of the Center, are complemented by program enrichments such as scholar exchanges and linkages to other NASA and commercial programs. The focus of this training program, and a preliminary assessment of its successes, are described.

Guikema, James A.↗

NASA NTRS1990

Binding of alpha-fetoprotein by immobilized monoclonal antibodies during episodes of zero-gravity obtained by parabolic flight

Alpha-fetoprotein (AFP), a single-chain polypeptide which is synthesized by the liver and yolk sac of the human fetus, provided a model ligand for assessing the effects of microgravity on ligand binding to surface-immobilized model receptor molecules. Monoclonal antibodies, used as receptors for AFP, were immobilized by covalent attachment to latex microparticles. Zero gravity environment was obtained by parabolic flight aboard NASA 930, a modified KC-135 aircraft. Buring the onset of an episode of zero gravity, ligand and receptor were mixed. Timed incubation (20 s) was terminated by centrifugation, the supernatant removed, and microparticies were assessed for bound AFP by immunochemical methods. The extent of binding was not influenced by microgravity, when compared with 1-G controls, which suggests that aberrant cellular activities observed in microgravity are not the simple expression of altered macromolecular interactions.

Spooner, Brian S.↗

NASA NTRS1990

Performance of a blood chemistry analyzer during parabolic flight

The performance of the Vision System Blood Analyzer during parabolic flight on a KC-135 aircraft (NASA 930) has been tested. This fully automated instrument performed flawlessly in these trials, demonstrating its potential for efficient, reliable use in a microgravity environment. In addition to instrument capability, it is demonstrated that investigators could readily fill specially modified test packs with fluid during zero gravity, and that filled test packs could be easily loaded into VISION during an episode of microgravity.

Spooner, Brian S.↗