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At least 181 records · Page 10

Cloning higher plants from aseptically cultured tissues and cells

A review of aseptic culture methods for higher plants is presented, which focuses on the existing problems that limit or prevent the full realization of cloning plants from free cells. It is shown that substantial progress in clonal multiplication has been made with explanted stem tips or lateral buds which can be stimulated to produce numerous precocious axillary branches. These branches can then be separated or subdivided and induced to root in order to yield populations of genetically and phenotypically uniorm plantlets. Similarly, undifferentiated calluses can sometimes be induced to form shoots and/or roots adventitiously. Although the cell culture techniques required to produce somatic embryos are presently rudimentary, steady advances are being made in learning how to stimulate formation of somatic or adventive embryos from totipotent cells grown in suspension cultures. It is concluded that many problems exist in the producing and growing of totipotent or morphogenetically competent cell suspensions, but the potential benefits are great.

Krikorian, A. D.↗

Regulatory coding of lymphoid lineage choice by hematopoietic transcription factors

During lymphopoiesis, precursor cells negotiate a complex regulatory space, defined by the levels of several competing and cross-regulating transcription factors, before arriving at stable states of commitment to the B-, T- and NK-specific developmental programs. Recent perturbation experiments provide evidence that this space has three major axes, corresponding to the PU.1 versus GATA-1 balance, the intensity of Notch signaling through the CSL pathway, and the ratio of E-box transcription factors to their Id protein antagonists.

Non-NASA Center↗

Hematological measurements in rats flown on Spacelab shuttle, SL-3

Previous studies have shown that a decrease in red cell mass occurs in astronauts, and some studies indicate a leukocytosis occurs. A life science module housing young and mature rats was flown on shuttle mission Spacelab 3 (SL-3), and the results of hematology studies of flight and control rats are presented. Statistically significant increases in the hematocrit, red blood cell counts, and hemoglobin determinations, together with a mild neutrophilia and lymphopenia, were found in flight animals. No significant changes were found in bone marrow and spleen cell differentials or erythropoietin determinations. Clonal assays demonstrated an increased erythroid colony formation of flight animal bone marrow cells at erythropoietin doses of 0.02 and 1.0 U/ml but not 0.20 U/ml. These results agree with some but vary from other previously published studies. Erythropoietin assays and clonal studies were performed for the first time.

NASA Experiment Number SL3-26↗

The structure of the stem endodermis in etiolated pea seedlings

Differentiation of the endodermis was examined in third internodes of etiolated Pisum sativum L. cv. Alaska seedlings. The endodermis in young internodes contains large, sedimented amyloplasts; in older internodes, a casparian strip differentiates and the endodermis becomes depleted of starch except for the proximal region of the stem, which retains sedimented amyloplasts and remains graviresponsive. Sedimentation occurs in the hook but does not occur consistently until cells reach the base of the hook, where the axis becomes vertical, rapid cell elongation starts, and amyloplast diameter increases substantially. Contact between endoplasmic reticulum and amyloplasts was observed. Endoplasmic reticulum is not distributed polarly with respect to gravity. No symplastic or apoplastic blockages exist in the endodermis at the level of the stem where lateral gradients may be established during tropic curvature.

Non-NASA Center↗

Progress Towards a Cartesian Cut-Cell Method for Viscous Compressible Flow

We present preliminary development of an approach for simulating high Reynolds number steady compressible flow in two space dimensions using a Cartesian cut-cell finite volume method. We consider both laminar and turbulent flow with both low and high cell Reynolds numbers near the wall. The approach solves the full Navier-Stokes equations in all cells, and uses a wall model to address the resolution requirements near boundaries and to mitigate mesh irregularities in cut cells. We present a quadratic wall model for low cell Reynolds numbers. At high cell Reynolds numbers, the quadratic is replaced with a newly developed analytic wall model stemming from solution of a limiting form of the Spalart-Allmaras turbulence model which features a forward evaluation for flow velocity and exactly matches characteristics of the SA turbulence model in the field. We develop multigrid operators which attain convergence rates similar to inviscid multigrid. Investigations focus on preliminary verification and validation of the method. Flows over flat plates and compressible airfoils show good agreement with both theoretical results and experimental data. Mesh convergence studies on sub- and transonic airfoil flows show convergence of surface pressures with wall spacings as large as approx.0.1% chord. With the current analytic wall model, one or two additional refinements near the wall are required to obtain mesh converged values of skin friction.

Berger, Marsha↗

Light Spectral Quality Effects on the Growth of Potato (Solanum Tuberosum L.) Nodal Cuttings in Vitro

The effects of light spectral quality on the growth of in vitro nodal cuttings of potato (Solanum tuberosum L.) cultivars Norland, Superior, Kennebec, and Denali were examined. The different light spectra were provided by Vita-Lite fluorescent (VF) (a white light control), blue fluorescent (BF), red fluorescent (RF), low-pressure sodium (LPS), and a combination of low-pressure sodium plus cool-white fluorescent lamps (LPS/CWF). For cultivars, stem lengths after 4 wks were longest under LPS, follow by RF, LPS/CWF, VF, and BF (in descending order). Microscopic studies revealed that cells were shortest when cultured in BF or VF environments, and were longest in RF or LPS lamp environments. The highest number axillary branches occurred on plantlets grown with LPS or LPS/CWF, whereas the lowest number occurred with BF. No leaf or stem edema (callus or gall-like growths) occurred iwth LPS or LPS/cwf lighting, and no edema occurred on cv. Norland plantlets, regardless of lighting. Results suggest that shoot morphologic development of in vitro grown potato plants can be controlled by controlling irradiant spectral quality.

Wilson, Deborah A.↗

Gravitropism in plants: Hydraulics and wall growth properties of responding cells

Gravitropism is the asymmetrical alteration of plant growth in response to a change in the gravity vector, with the typical result that stems grow up and roots grow down. The gravity response is important for plants because it enables them to grow their aerial parts in a mechanically stable (upright) position and to develop their roots and leaves to make efficient use of soil nutrients and sunlight. The elucidation of gravitropic responses will tell much about how gravity exerts its morphogenetic effects on plants and how plants regulate their growth at the cellular and molecular levels.

Cosgrove, Daniel J.↗

Electromagnetically Clean Solar Arrays

The term 'electromagnetically clean solar array' ('EMCSA') refers to a panel that contains a planar array of solar photovoltaic cells and that, in comparison with a functionally equivalent solar-array panel of a type heretofore used on spacecraft, (1) exhibits less electromagnetic interferences to and from other nearby electrical and electronic equipment and (2) can be manufactured at lower cost. The reduction of electromagnetic interferences is effected through a combination of (1) electrically conductive, electrically grounded shielding and (2) reduction of areas of current loops (in order to reduce magnetic moments). The reduction of cost is effected by designing the array to be fabricated as a more nearly unitary structure, using fewer components and fewer process steps. Although EMCSAs were conceived primarily for use on spacecraft they are also potentially advantageous for terrestrial applications in which there are requirements to limit electromagnetic interference. In a conventional solar panel of the type meant to be supplanted by an EMCSA panel, the wiring is normally located on the back side, separated from the cells, thereby giving rise to current loops having significant areas and, consequently, significant magnetic moments. Current-loop geometries are chosen in an effort to balance opposing magnetic moments to limit far-0field magnetic interactions, but the relatively large distances separating current loops makes full cancellation of magnetic fields problematic. The panel is assembled from bare photovoltaic cells by means of multiple sensitive process steps that contribute significantly to cost, especially if electomagnetic cleanliness is desired. The steps include applying a cover glass and electrical-interconnect-cell (CIC) sub-assemble, connecting the CIC subassemblies into strings of series-connected cells, laying down and adhesively bonding the strings onto a panel structure that has been made in a separate multi-step process, and mounting the wiring on the back of the panel. Each step increases the potential for occurrence of latent defects, loss of process control, and attrition of components. An EMCSA panel includes an integral cover made from a transparent material. The silicone cover supplants the individual cover glasses on the cells and serves as an additional unitary structural support that offers the advantage, relative to glass, of the robust, forgiving nature of the silcone material. The cover contains pockets that hold the solar cells in place during the lamination process. The cover is coated with indium tin oxide to make its surface electrically conductive, so that it serves as a contiguous, electrically grounded shield over the entire panel surface. The cells are mounted in proximity to metallic printed wiring. The painted-wiring layer comprises metal-film traces on a sheet of Kapton (or equivalent) polyimide. The traces include contact pads on one side of the sheet for interconnecting the cells. Return leads are on the opposite side of the sheet, positioned to form the return currents substantially as mirror images of, and in proximity to, the cell sheet currents, thereby minimizing magnetic moments. The printed-wiring arrangement mimics the back-wiring arrangement of conventional solar arrays, but the current-loop areas and the resulting magnetic moments are much smaller because the return-current paths are much closer to the solar-cell sheet currents. The contact pads are prepared with solder fo electrical and mechanical bonding to the cells. The pocketed cover/shield, the solar cells, the printed-wiring layer, an electrical bonding agent, a mechanical-bonding agent, a composite structural front-side face sheet, an aluminum honeycomb core, and a composite back-side face sheet are all assembled, then contact pads are soldered to the cells and the agents are cured in a single lamination process.

Stem, Theodore G.↗

Induced compression wood formation in Douglas fir (Pseudotsuga menziesii) in microgravity

In the microgravity environment of the Space Shuttle Columbia (Life and Microgravity Mission STS-78), were grown 1-year-old Douglas fir and loblolly pine plants in a NASA plant growth facility. Several plants were harnessed (at 45 degrees ) to establish if compression wood biosynthesis, involving altered cellulose and lignin deposition and cell wall structure would occur under those conditions of induced mechanical stress. Selected plants were harnessed at day 2 in orbit, with stem sections of specific plants harvested and fixed for subsequent microscopic analyses on days 8, 10 and 15. At the end of the total space mission period (17 days), the remaining healthy harnessed plants and their vertical (upright) controls were harvested and fixed on earth. All harnessed (at 45 degrees ) plant specimens, whether grown at 1 g or in microgravity, formed compression wood. Moreover, not only the cambial cells but also the developing tracheid cells underwent significant morphological changes. This indicated that the developing tracheids from the primary cell wall expansion stage to the fully lignified maturation stage are involved in the perception and transduction of the stimuli stipulating the need for alteration of cell wall architecture. It is thus apparent that, even in a microgravity environment, woody plants can make appropriate corrections to compensate for stress gradients introduced by mechanical bending, thereby enabling compression wood to be formed. The evolutionary implications of these findings are discussed in terms of "variability" in cell wall biosynthesis.

manned↗

Descending pathways to the cutaneus trunci muscle motoneuronal cell group in the cat

The descending pathways to the motoneuronal cell group of the cutaneous trunci muscle (CTM) of the cat were investigated by injecting H-3-labeled lucine into the brain stem, the diencephalon, or the C1, C2, C6, and C8 segments of the spinal cord, and examining fixed autoradiographic sections of the spinal cord and brain regions. Results demonstrate presence of specific supraspinal projectons to the CTM motor nucleus originating in the contralateral nucleus retroambiguous and the ipsilateral dorsolateral pontine tegmentum. Results also suggest that propriospinal pathways to the CTM motor nucleus originating in the cervical cord do not exist, although these propriospinal projections to all other motoneuronal cell groups surrounding the CTM nucleus are very strong.

Holstege, Gert↗

The role of cell membranes in the regulation of lignification in pine cells

The identity of pine cell membranes bearing PAL enzyme activity, the isolation of a plasma membrane preparation from pine cells for testing as a regulatory barrier in lignification, and the measurement of the geopotential effect in pine stems are presented. A model to describe and predict the interaction of gravity and lignification of higher plants was developed.

Hendrix, D. L.↗

Isolation and characterization of beta-glucan synthase: A potential biochemical regulator of gravistimulated differential cell wall loosening

In plants, gravity stimulates differential growth in the upper and lower halves of horizontally oriented organs. Auxin regulation of cell wall loosening and elongation is the basis for most models of this phenomenon. Auxin treatment of pea stem tissue rapidly increases the activity of Golgi-localized Beta-1,4-glucan synthase, an enzyme involved in biosynthesis of wall xyloglucan which apparently constitutes the substrate for the wall loosening process. The primary objective is to determine if auxin induces de novo formation of Golgi glucan synthase and increases the level of this glucan synthase mRNA. This shall be accomplished by (a) preparation of a monoclonal antibody to the synthase, (b) isolation, and characterization of the glucan synthase, and (c) examination for cross reactivity between the antibody and translation products of auxin induced mRNAs in pea tissue. The antibody will also be used to localize the glucan synthase in upper and lower halves of pea stem tissue before, during and after the response to gravity.

Kuzmanoff, K. M.↗

Isolation of Hybridomas for Golgi-associated Proteins and a Plant Calmodulin

The demonstration of a role for calcium in the mechanism of the gravitropic response indicates a role for calmodulin. Localization studies indicate that plant cell walls have a high content of calmodulin which suggests a regulatory role for CaM in both gravitropic curvature and auxin-induced growth. Auxin regulation of cell wall loosening and elongation is the basis for most models of this phenomenon. Auxin treatment of pea stem tissue rapidly increases the ctivity of Golgi-localized B-1,4-glucan synthase (GS), an enzyme involved in biosynthesis of wall xyloglucan which apparently constitutes the substrate for the wall loosening process. In order to determine whether auxin stimulates GS activity either by modulation of existing enzyme or induces de novo formation of Golgi glucan synthase, a study was undertaken to isolate and quantitate glucan synthase. This enzyme appears to be an integral protein of the Golgi membrane and has resisted isolation with retention of activity. The production of monoclonal antibody for glucan synthase was undertaken due to the inability to isolate GS by standard detergent/liposome techniques.

Kuzmanoff, K. M.↗

Effect of Microgravity on Afferent Innervation

Presentations and publications are: (1) an audiovisual summary web presentation on results from SLM-MIR avian experiments. A color presentation summarizing results from the SLM-MIR and STS-29 avian experiments; (2) color threshold and ratio of S 100B MAP5, NF68/200, GABA and GAD; (3) chicken (Gallus domesticus) inner ear afferents; (4) microgravity in the STS-29 Space Shuttle Discovery affected the vestibular system of chick embryos; (5) expression of S 100B in sensory and secretory cells of the vertebrate inner ear; (6) otoconia biogenesis, phylogeny, composition and functional attributes;(7) the glycan keratin sulfate in inner ear crystals; (8) elliptical-P cells in the avian perilymphatic interface of the tegmentum vasculosum; and (9) LAMP2c and S100B upregulation in brain stem after VIIIth nerve deafferentation.

Source record↗

Descending pathways to the cutaneus trunci muscle motoneuronal cell group in the cat

Pathways involved in the cutaneous trunci muscle (CTM) reflex in the cat were investigated. Experimental animals were injected with tritium-labeled L-leucine into their spinal cord, brain stem, or diencephalon and, after six weeks, perfused with 10-percent formalin. The brains and spinal cords were postfixed in formalin and were cut into transverse 25-micron-thick frozen sections for autoradiography. Results based on injections in the C1, C2, C6, and C8 segments suggest that propriospinal pathways to the CTM motor nucleus originating in the cervical cord do no exist, although these propriospinal projections are very strong to all other motoneuronal cell groups surrounding the CTM motor nucleus. The results also demonstrate presence of specific supraspinal projections to the CTM motor nucleus, originating in the contralateral nucleus retroambiguous and the ipsilateral dorsolateral pontine tegmentum.

Holstege, Gert↗

Localization and pattern of graviresponse across the pulvinus of barley Hordeum vulgare

Pulvini of excised stem segments from barley (Hordeum vulgare cv Larker') were pretreated with 1 millimolar coumarin before gravistimulation to reduce longitudinal cell expansion and exaggerate radial cell enlargement. The cellular localization and pattern of graviresponse across individual pulvini were then evaluated by cutting the organ in cross-section, photographing the cross-section, and then measuring pulvinus thickness and the radial width of cortical and epidermal cells in enlargements of the photomicrographs. With respect to orientation during gravistimulation, we designated the uppermost point of the cross-section 0 degrees and the lowermost point 180 degrees. A gravity-induced increase in pulvinus thickness was observable within 40 degrees of the vertical in coumarin-treated pulvini. In upper halves of coumarin-treated gravistimulated pulvini, cells in the inner cortex and inner epidermis had increased radial widths, relative to untreated gravistimulated pulvini. In lower halves of coumarin-treated pulvini, cells in the central and outer cortex and in the outer epidermis showed the greatest increase in radial width. Cells comprising the vascular bundles also increased in radial width, with this pattern following that of the central cortex. These results indicate (a) that all cell types are capable of showing a graviresponse, (b) that the graviresponse occurs in both the top and the bottom of the responding organ, and (c) that the magnitude of the response increases approximately linearly from the uppermost point to the lowermost. These results are also consistent with models of gravitropism that link the pattern and magnitude of the graviresponse to graviperception via statolith sedimentation.

Non-NASA Center↗

Re-Evaluation of the Role of Starch in Gravitropic Sensing

Plant organs grow toward or away from gravity as a way to orient those organs for optimizing growth. Starch has long been thought to be important in sensing the direction of the g-vector in gravitropism, but that hypothesis has also evoked controversy. We have previously shown that starch-deficient mutants of Arabidopsis (TC7) and Nicotiana (NS458) are impaired in their gravitropism. While this suggests that starch is not necessary for reduced gravitropism, it also indicates that the mass of the starch contributes to sensing when present and thus is necessary for full gravitropic sensitivity. The research supported by this grant focused on three related projects, (1) the effect of light on hypocotyl gravitropism in NS458, (2) the effects of root phototropism on measurements of gravitropic sensitivity, and (3) the effects of starch overproduction on sedimentation and gravitropism. Collectively, our results provide additional strong support for the importance of starch in gravitropic sensing. First, by accounting for negative phototropism in roots of two starchless mutants of Arabidopsis we have established that these mutants are much less sensitive to gravity than previously thought. This work also demonstrates the importance of designing experimental protocols that remove the influence of root phototropism on measuring root gravitropism. Second, light apparently promotes gravitropism in starch-deficient Nicotiana hypocotyls by increasing the trace amounts of starch in the plastids, by inducing limited plastid sedimentation and thus by presumably increasing the signal provided by plastid mass. And finally, we show that excess starch in Arabidopsis seedlings has little effect on gravitropic sensitivity implying that the sensing system is already saturated. However, in light-grown stems where this mutation results in starch accumulation and where the wild-type practically lacks starch in the sensing cells, the mutant is much more sensitive than the wild-type again showing that the loss of starch depresses gravity sensing.

Sack, Fred D.↗

Differential expression of members of the annexin multigene family in Arabidopsis

Although in most plant species no more than two annexin genes have been reported to date, seven annexin homologs have been identified in Arabidopsis, Annexin Arabidopsis 1-7 (AnnAt1--AnnAt7). This establishes that annexins can be a diverse, multigene protein family in a single plant species. Here we compare and analyze these seven annexin gene sequences and present the in situ RNA localization patterns of two of these genes, AnnAt1 and AnnAt2, during different stages of Arabidopsis development. Sequence analysis of AnnAt1--AnnAt7 reveals that they contain the characteristic four structural repeats including the more highly conserved 17-amino acid endonexin fold region found in vertebrate annexins. Alignment comparisons show that there are differences within the repeat regions that may have functional importance. To assess the relative level of expression in various tissues, reverse transcription-PCR was carried out using gene-specific primers for each of the Arabidopsis annexin genes. In addition, northern blot analysis using gene-specific probes indicates differences in AnnAt1 and AnnAt2 expression levels in different tissues. AnnAt1 is expressed in all tissues examined and is most abundant in stems, whereas AnnAt2 is expressed mainly in root tissue and to a lesser extent in stems and flowers. In situ RNA localization demonstrates that these two annexin genes display developmentally regulated tissue-specific and cell-specific expression patterns. These patterns are both distinct and overlapping. The developmental expression patterns for both annexins provide further support for the hypothesis that annexins are involved in the Golgi-mediated secretion of polysaccharides.

Non-NASA Center↗