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At least 217 records · Page 12

Root gravitropism in response to a signal originating outside of the cap

We have developed image analysis software linked to a rotating stage, allowing constraint of any user-selected region of a root at a prescribed angle during root gravitropism. This device allows the cap of a graviresponding root to reach vertical while maintaining a selected region within the elongation zone at a gravistimulated angle. Under these conditions gravitropic curvature of roots of Zea mays L. continues long after the root cap reaches vertical, indicating that a signal from outside of the cap can contribute to the curvature response.

Non-NASA Center↗

Early root cap development and graviresponse in white clover (Trifolium repens) grown in space and on a two-axis clinostat

White clover (Trifolium repens) was germinated and grown in microgravity aboard the Space Shuttle (STS-60, 1994; STS-63, 1995), on Earth in stationary racks and in a slow-rotating two-axis clinostat. The objective of this study was to determine if normal root cap development and early plant gravity responses were dependent on gravitational cues. Seedlings were germinated in space and chemically fixed in orbit after 21, 40, and 72 h. Seedlings 96 h old were returned viable to earth. Germination and total seedling length were not dependent on gravity treatment. In space-flown seedlings, the number of cell stories in the root cap and the geometry of central columella cells did not differ from those of the Earth-grown seedlings. The root cap structure of clinorotated plants appeared similar to that of seedlings from microgravity, with the exception of three-day rotated plants, which displayed significant cellular damage in the columella region. Nuclear polarity did not depend on gravity; however, the positions of amyloplasts in the central columella cells were dependent on both the gravity treatment and the age of the seedlings. Seedlings from space, returned viable to earth, responded to horizontal stimulation as did 1 g controls, but seedlings rotated on the clinostat for the same duration had a reduced curvature response. This study demonstrates that initial root cap development is insensitive to either chronic clinorotation or microgravity. Soon after differentiation, however, clinorotation leads to loss of normal root cap structure and plant graviresponse while microgravity does not.

manned↗

A morphometric analysis of the redistribution of organelles in columella cells of horizontally-oriented roots of Zea mays

In order to determine what structural changes in graviperceptive cells are associated with onset of root gravicurvature, the redistribution of organelles in columella cells of horizontally-oriented, graviresponding roots of Zea mays has been quantified. Root gravicurvature began by 15 min after reorientation, and did not involve significant changes in the (i) volume of individual columella cells or amyloplasts, (ii) relative volume of any cellular organelle, (iii) number of amyloplasts per columella cell, or (iv) surface area of cellular location of endoplasmic reticulum. Sedimentation of amyloplasts began within 1 to 2 min after reorientation, and was characterized by an intensely staining area of cytoplasm adjacent to the sedimenting amyloplasts. By 5 min after reorientation, amyloplasts were located in the lower distal corner of columella cells, and, by 15 min after reorientation, overlaid the entire length of the lower cell wall. No consistent contact between amyloplasts and any cellular structure was detected at any stage of gravicurvature. Centrally-located nuclei initially migrated upward in columella cells of horizontally-oriented roots, after which they moved to the proximal ends of the cells by 15 min after reorientation. No significant pattern of redistribution of vacuoles, mitochondria, dictyosomes, or hyaloplasm was detected that correlated with the onset of gravicurvature. These results indicate that amyloplasts and nuclei are the only organelles whose movements correlate positively with the onset of gravicurvature by primary roots of this cultivar of Zea mays.

NASA Discipline Plant Biology↗

How roots perceive and respond to gravity

Graviperception by plant roots is believed to occur via the sedimentation of amyloplasts in columella cells of the root cap. This physical stimulus results in an accumulation of calcium on the lower side of the cap, which in turn induces gravicurvature. In this paper we present a model for root gravitropism integrating gravity-induced changes in electrical potential, cytochemical localization of calcium in cells of gravistimulated roots, and the interdependence of calcium and auxin movement. Key features of the model are that 1) gravity-induced redistribution of calcium is an early event in the transduction mechanism, and 2) apoplastic movement of calcium through the root-cap mucilage may be an important component of the pathway for calcium movement.

Non-NASA Center↗

Light-regulated protein and mRNA synthesis in root caps of maize

Illumination of maize roots initiates changes in mRNA levels and in the activities of proteins within the root cap. Using Northern analysis we showed a 5-6 fold increase in the levels of three specific mRNAs and a 14-fold increase in plastid mRNA. This increase is rapid, occurring within 30 minutes of illumination. With prolonged periods of darkness following illumination, messages return to levels observed in dark, control caps. For two species of mRNA illumination results in a reduction in message levels. Light-stimulated increases in the levels of specific mRNAs are proportionally greater than are increases in the activities of corresponding proteins. We suggest that the light-stimulated increase in protein activity in root caps may be preceded by and occur as a consequence of enhanced levels of mRNA. Our work suggests that photomorphogenesis in roots could involve changes in the levels of a wide variety of mRNAs within the root cap.

NASA Program Space Biology↗

Microsurgical removal of epidermal and cortical cells: evidence that the gravitropic signal moves through the outer cell layers in primary roots of maize

There is general agreement that during root gravitropism some sort of growth-modifying signal moves from the cap to the elongation zone and that this signal ultimately induces the curvature that leads to reorientation of the root. However, there is disagreement regarding both the nature of the signal and the pathway of its movement from the root cap to the elongation zone. We examined the pathway of movement by testing gravitropism in primary roots of maize (Zea mays L.) from which narrow (0.5 mm) rings of epidermal and cortical tissue were surgically removed from various positions within the elongation zone. When roots were girdled in the apical part of the elongation zone gravitropic curvature occurred apical to the girdle but not basal to the girdle. Filling the girdle with agar allowed curvature basal to the girdle to occur. Shallow girdles, in which only two or three cell layers (epidermis plus one or two cortical cell layers) were removed, prevented or greatly delayed gravitropic curvature basal to the girdle. The results indicate that the gravitropic signal moves basipetally through the outermost cell layers, perhaps through the epidermis itself.

NASA Discipline Number 29-20↗

Chapter 7: Fluorescence Imaging of the Cytoskeleton in Plant Roots

During the past two decades the use of live cytoskeletal probes has increased dramatically due to the introduction of the green fluorescent protein. However, to make full use of these live cell reporters it is necessary to implement simple methods to maintain plant specimens in optimal growing conditions during imaging. To image the cytoskeleton in living Arabidopsis roots, we rely on a system involving coverslips coated with nutrient supplemented agar where the seeds are directly germinated. This coverslip system can be conveniently transferred to the stage of a confocal microscope with minimal disturbance to the growth of the seedling. For roots with a larger diameter such as Medicago truncatula, seeds are first germinated in moist paper, grown vertically in between plastic trays, and roots mounted on glass slides for confocal imaging. Parallel with our live cell imaging approaches, we routinely process fixed plant material via indirect immunofluorescence. For these methods we typically use non-embedded vibratome-sectioned and whole mount permeabilized root tissue. The clearly defined developmental regions of the root provide us with an elegant system to further understand the cytoskeletal basis of plant development.

Actin↗

Root Dynamics Mitigate Warm and Dry Biases over the Central United States

The central United States frequently exhibits warm and dry biases in simulations of summertime conditions, a persistent feature that remains unresolved. While previous studies linked these biases to misrepresented surface energy exchanges, the role of belowground processes remains poorly understood. Here, we demonstrate that inadequate representation of root water uptake in land surface models contributes to this bias. Using both offline Noah-MP and coupled WRF-Noah-MP simulations with static and dynamic root water uptake schemes, we show that the inclusion of dynamic root processes reduces 2-m air temperature biases and enhances precipitation, primarily by increasing the rain rate of convective systems. Offline and coupled simulations further reveal that the cooling effects and precipitation increases are amplified through positive land-atmosphere feedback, active only in the coupled model. These findings highlight an important role of root in modulating land-atmosphere interactions and underscore the need to refine root-zone processes to improve regional atmospheric simulations.

Yang, Zhao (ORCID:0000000288027130)↗

Enrichment of root-associated Streptomyces strains in response to drought is driven by diverse functional traits and does not predict beneficial effects on plant growth

The genus Streptomyces has consistently been found enriched in drought-stressed plant root microbiomes, yet the ecological basis and functional variation underlying this enrichment at the strain and isolate level remain unclear. Using two 16S rRNA sequencing methods with different levels of taxonomic resolution, we confirmed drought-associated enrichment (DE) of Streptomyces in field-grown sorghum roots and identified five closely related but distinct amplicon sequence variants (ASVs) belonging to the genus with variable drought enrichment patterns. From a culture collection of sorghum root endophytes, we selected 12 Streptomyces isolates representing these ASVs for phenotypic and genomic characterization. Whole-genome sequencing revealed substantial variation in gene content, even among closely related isolates, and exometabolomic profiling showed distinct metabolic responses to media supplemented with drought- versus well-watered root tissue. Traits linked to drought survival, including osmotic stress tolerance, siderophore production, and carbon utilization, varied widely among isolates and were not phylogenetically conserved. Using a broader panel of 48 Streptomyces, we demonstrate that DE scores, determined through mono-association experiments in gnotobiotic sorghum systems, showed high variability and lacked correlation with plant growth promotion. Pangenome-wide association identified orthogroups involved in osmolyte transport (e.g., proP) and membrane biosynthesis (e.g., fabG) as positively associated with DE, though most associations lacked phylogenetic signal. Collectively, these results demonstrate that Streptomyces DE is not a conserved genus-level trait but is instead strain-specific and functionally heterogeneous. Furthermore, DE in the root microbiome was shown not to predict beneficial effects on plant growth. This work underscores the need to resolve functional traits at the strain level and highlights the complexity of microbe-host-environment interactions under abiotic stress.

Fonseca-Garcia, Citlali↗

Accelerating the identification of novel secondary metabolites in bioenergy plant root exudates using MicroED

Small molecule metabolites drive inter- and intraspecies communication and dependencies in diverse biological systems, yet a large proportion of these important chemical compounds remain uncharacterized in plants and microbes. Approximately 90% of the metabolites in root exudate profiles are unknown compounds, despite the importance of root exudate composition in plant-microbe interactions. We need advanced analytical capabilities that will support rapid discovery and structural elucidation of metabolites from biological samples that may be limited in quantity and high in complexity. To fill this gap, this project aimed to develop an integrated workflow involving metabolite extraction, separation, and crystallization from plant root exudates followed by characterization using nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry, and microcrystal electron diffraction (MicroED). Using crude root exudates from sorghum, this project successfully developed higher throughput exudate fractionation strategies to obtain pure compounds for crystallization and identified crystals in multiple fractions that diffracted. Additional efforts to increase the throughput of high-quality crystal generation for MicroED, such as crystallization screening and crystallization chaperone exploration, will be needed to further advance root exudate metabolite identification. The overall optimized sample preparation process can then be integrated with the existing data collection and data analysis pipelines for MicroED at PNNL to facilitate more rapid natural product discovery.

59 BASIC BIOLOGICAL SCIENCES↗

Roots of polynomials by ratio of successive derivatives

An order of magnitude study of the ratios of successive polynomial derivatives yields information about the number of roots at an approached root point and the approximate location of a root point from a nearby point. The location approximation improves as a root is approached, so a powerful convergence procedure becomes available. These principles are developed into a computer program which finds the roots of polynomials with real number coefficients.

Crouse, J. E.↗

Effect of wing sweep, angle of attack, Reynolds number, and wing root fillet on the interference heating to the wing windward surface of an entry vehicle configuration

The phase-change-coating technique was used to study the interference heating to the windward surface of 14 deg, 25 deg, and 50 deg swept wings of an entry vehicle configuration. One wing root of each model was faired to the fuselage with a fillet. Tests were made at Mach 8 at angles of attack of 0 deg, 20 deg, 40 deg, and 60 deg and at free-stream Reynolds numbers based on model length of 0.47 and 1.7 million. Bow shock impingement heating was found to increase in magnitude and affected area with increasing angle of attack until at a higher angle of attack it decreases; this angle of attack is lower for a 50 deg swept wing. Wing root interference heating was found to increase with angle of attack up to 40 deg and then to remain approximately constant. Consequently, wing root interference heating becomes the major type of interference heating at large angles of attack, and this occurs at a lower angle of attack for the highest sweep angle. A wing leading-edge root fillet reduces the peak in wing root interference heating near the leading edge, and increasing Reynolds number increases the level of interference heating.

Clark, L. E.↗

Protein synthesis in geostimulated root caps

A study is presented of the processes occurring in the root cap of corn which are requisite for the formation of root cap inhibitor and which can be triggered or modulated by both light and gravity. The results of this study indicate the importance of protein synthesis for light-induced gravitropic bending in roots. Root caps in which protein synthesis is prevented are unable to induce downward bending. This suggests that light acts by stimulating proteins which are necessary for the translation of the gravitropic stimulus into a growth response (downward bending). The turnover of protein with time was also examined in order to determine whether light acts by stimulating the synthesis of unique proteins required for downward growth. It is found that auxin in combination with light allows for the translation of the gravitropic stimulus into a growth response at least in part through the modification of protein synthesis. It is concluded that unique proteins are stimulated by light and are involved in promoting the downward growth in roots which are responding to gravity.

Feldman, L. J.↗

A comparative study of monocot and dicot root development in normal /earth/ and hypogravity /space/ environments

The anatomy and fine structure of roots from oat and mung bean seedlings grown under hypogravity conditions aboard NASA's Space Shuttle were examined and compared to those of roots from ground control plants grown under similar conditions. Oat roots from both sets of plants exhibited normal tissue organization and ultrastructural features, with the exception of cortex cell mitochondria, which characteristically showed a 'swollen' morphology. Flight-grown mung bean roots differed significantly from the controls in that root cap cells were somewhat disorganized and degraded in appearance, especially at the cap periphery. At the EM level, these cells exhibited a loss organelle integrity and a condensed cytoplasm. The potential significance of this finding for the putative gravity-sensing cap cells were noted.

Slocum, R. D.↗

Cytochemical localization of calcium in cap cells of primary roots of Zea mays L

The cellular distribution of Ca in caps of primary roots of Zea mays was examined during the onset and early stages of gravicurvature to determine its possible role in root gravitropism. Staining becomes associated with the portion of the cell wall adjacent to the distal end of the cell after five minutes, and persists throughout the onset of gravicurvature. The outermost peripheral cells of roots oriented horizontally and vertically secrete Ca through plasmodesmata-like channels in their cell walls. Data suggest that Ca is not transported laterally through the columella tissue,but rather that the movement of Ca to the lower side of caps of horizontally-oriented roots is at least partially through and/or on the mucilage of the cap, and via an electrochemical gradient. An important role in root gravitropism is indicated for Ca secretion by peripheral cells.

Moore, R.↗

The role of calcium and calmodulin in the response of roots to gravity

There is general agreement that, in roots, the primary detection of the gravitropic signal occurs in the columella cells of the cap and that this results in the generation of a signal that moves into the elongation zone causing the asymmetric growth that leads to downward curvature. Recent work has generated considerable evidence that indicates that auxin is the ultimate mediator of differential growth during root (and shoot) gravitropism. Our studies of the time course of curvature, auxin redistribution and/or adaptation, and electrical potential changes in maize roots have led to the following generalizations: (1) downward curvature begins 18 to 32 min following gravistimulation; (2) asymmetric auxin redistribution across the root cap begins at about the same time as curvature or perhaps slight earlier; (3) there is a lag of approx. 15 min in the response of roots to applied auxin; and (4) gravi-induced changes in intracellular potentials of cortical cells within the elongation zone occur within 30 s following stimulation.

Evans, Michael L.↗

Locating three-dimensional roots by a bisection method

Bisection methods, which depend on the existence of a criterion for determining whether a root exists within a given volume, may be useful in the evaluation of equation roots since the volume in which the root is known to be located can be steadily decreased. This criterion is presently furnished via topological degree theory, strictly for the case of 3D volumes; usefulness is demonstrated in the location of roots, and the classification of roots as either X- or O-points is discussed.

Greene, John M.↗

Root growth and development in response to CO2 enrichment

A non-destructive technique (minirhizotron observation tubes) was used to assess the effects of CO2 enrichment on root growth and development in experimental plots in a scrub oak-palmetto community at the Kennedy Space Center. Potential effects of CO2 enrichment on plants have a global significance in light of concerns over increasing CO2 concentrations in the Earth's atmosphere. The study at Kennedy Space Center focused on aboveground physiological responses (photosynthetic efficiency and water use efficiency), effects on process rates (litter decomposition and nutrient turnover), and belowground responses of the plants. Belowground dynamics are an exceptionally important component of total plant response but are frequently ignored due to methodological difficulties. Most methods used to examine root growth and development are destructive and, therefore, severely compromise results. Minirhizotrons allow nondestructive observation and quantification of the same soil volume and roots through time. Root length density and root phenology were evaluated for CO2 effects with this nondestructive technique.

Day, Frank P., Jr.↗