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At least 73 records · Page 4

On the role of calcium in indole-3-acetic acid movement and graviresponse in etiolated pea epicotyls

To determine whether Ca2+ plays a special role in the early graviresponse of shoots, as has been reported for roots, we treated etiolated pea epicotyls with substances known to antagonize Ca2+ (La3+), to remove Ca2+ from the wall (spermidine, EGTA), to inhibit calmodulin mediated reactions (chlorpromazine), or to inhibit IAA transport (TIBA). We studied the effect of these substances on IAA and Ca2+ uptake into 7 mm long subapical 3rd internode etiolated pea epicotyl sections and pea leaf protoplasts, on pea epicotyl growth, and graviresponse and on lateral IAA redistribution during gravistimulation. Our results support the view that adequate Ca2+ in the apoplast is required for normal IAA uptake, transport and graviresponse. Experiments with protoplasts indicate that Ca2+ may be controlling a labile membrane porter, possibly located on the external surface of cell membrane, while inhibitor experiments suggest that calmodulin is also implicated in both the movement of IAA and graviresponse. Since a major transfer of Ca2+ through free space during graviresponse has not yet been demonstrated, and since inhibition of calcium channels does not affect IAA redistribution (Migliaccio and Galston, 1987, Plant Physiology 85:542), we conclude that no clear evidence links prior Ca2+ movement with IAA redistribution during graviresponse in stems.

Non-NASA Center↗

Amyloplasts are necessary for full gravitropic sensitivity in roots of Arabidopsis thaliana

The observation that a starchless mutant (TC7) of Arabidopsis thaliana (L.) Heynh. is gravitropic (T. Caspar and B.G. Pickard, 1989, Planta 177, 185-197) raises questions about the hypothesis that starch and amyloplasts play a role in gravity perception. We compared the kinetics of gravitropism in this starchless mutant and the wild-type (WT). Wild-type roots are more responsive to gravity than TC7 roots as judged by several parameters: (1) Vertically grown TC7 roots were not as oriented with respect to the gravity vector as WT roots. (2) In the time course of curvature after gravistimulation, curvature in TC7 roots was delayed and reduced compared to WT roots. (3) TC7 roots curved less than WT roots following a single, short (induction) period of gravistimulation, and WT, but not TC7, roots curved in response to a 1-min period of horizontal exposure. (4) Wild-type roots curved much more than TC7 roots in response to intermittent stimulation (repeated short periods of horizontal exposure); WT roots curved in response to 10 s of stimulation or less, but TC7 roots required 2 min of stimulation to produce a curvature. The growth rates were equal for both genotypes. We conclude that WT roots are more sensitive to gravity than TC7 roots. Starch is not required for gravity perception in TC7 roots, but is necessary for full sensitivity; thus it is likely that amyloplasts function as statoliths in WT Arabidopsis roots. Furthermore, since centrifugation studies using low gravitational forces indicated that starchless plastids are relatively dense and are the most movable component in TC7 columella cells, the starchless plastids may also function as statoliths.

NASA Discipline Plant Biology↗

Gravitropism in a starchless mutant of Arabidopsis: implications for the starch-statolith theory of gravity sensing

The starch-statolith theory of gravity reception has been tested with a mutant of Arabidopsis thaliana (L.) Heynh. which, lacking plastid phosphoglucomutase (EC 2.7.5.1) activity, does not synthesize starch. The hypocotyls and seedling roots of the mutant were examined by light and electron microscopy to confirm that they did not contain starch. In upright wild-type (WT) seedlings, starch-filled plastids in the starch sheath of the hypocotyl and in three of the five columellar layers of the root cap were piled on the cell floors, and sedimented to the ceilings when the plants were inverted. However, starchless plastids of the mutant were not significantly sedimented in these cells in either upright or inverted seedlings. Gravitropism of light-grown seedling roots was vigorous: e.g., 10 degrees curvature developed in mutants rotated on a clinostat following a 5 min induction at 1 g, compared with 14 degrees in the WT. Curvatures induced during intervals from 2.5 to 30 min were 70% as great in the mutant as the WT. Thus under these conditions the presence of starch and the sedimentation of plastids are unnecessary for reception of gravity by Arabidopsis roots. Gravitropism by hypocotyls of light-grown seedlings was less vigorous than that by roots, but the mutant hypocotyls exhibited an average of 70-80% as much curvature as the WT. Roots and hypocotyls of etiolated seedlings and flower stalks of mature plants were also gravitropic, although in these cases the mutant was generally less closely comparable to the WT. Thus, starch is also unnecessary for gravity reception in these tissues.

NASA Program Space Biology↗

Analysis of growth patterns during gravitropic curvature in roots of Zea mays by use of a computer-based video digitizer

A computer-based video digitizer system is described which allows automated tracking of markers placed on a plant surface. The system uses customized software to calculate relative growth rates at selected positions along the plant surface and to determine rates of gravitropic curvature based on the changing pattern of distribution of the surface markers. The system was used to study the time course of gravitropic curvature and changes in relative growth rate along the upper and lower surface of horizontally-oriented roots of maize (Zea mays L.). The growing region of the root was found to extend from about 1 mm behind the tip to approximately 6 mm behind the tip. In vertically-oriented roots the relative growth rate was maximal at about 2.5 mm behind the tip and declined smoothly on either side of the maximum. Curvature was initiated approximately 30 min after horizontal orientation with maximal (50 degrees) curvature being attained in 3 h. Analysis of surface extension patterns during the response indicated that curvature results from a reduction in growth rate along both the upper and lower surfaces with stronger reduction along the lower surface.

NASA Discipline Plant Biology↗

The role of calcium ions in gravity signal perception and transduction

Ca2+ is implicated as a messenger in coupling various environmental stimuli, such as gravity and light, to response. In recent years, it has become evident that Ca2+ plays a central role in all three phases of gravitropism--perception, transduction and response. The root cap, which is known to contain high amounts of Ca2+ and calmodulin, is the primary site of gravity perception. The possible role of phosphoinositide turnover and Ca(2+) - and Ca(2+) -calmodulin-dependent enzymes such as Ca(2+) -ATPase and protein kinases in gravitropism is discussed. A model is proposed to describe the role of Ca2+ in both normal and light-dependent gravity response in roots.

NASA Program Space Biology↗

The locations and amounts of endogenous ions and elements in the cap and elongating zone of horizontally oriented roots of Zea mays L.: an electron-probe EDS study

We used quantitative electron-probe energy-dispersive x-ray microanalysis to localize endogenous Na, Cl, K, P, S, Mg and Ca in cryofixed and freeze-dried cryosections of the cap (i.e. the putative site of graviperception) and elongating zone (i.e. site of gravicurvature) of horizontally oriented roots of Zea mays. Ca, Na, Cl, K and Mg accumulate along the lower side of caps of horizontally oriented roots. The most dramatic asymmetries of these ions occur in the apoplast, especially the mucilage. We could not detect any significant differences in the concentrations of these ions in the central cytoplasm of columella cells along the upper and lower sides of caps of horizontally-oriented roots. However, the increased amounts of Na, Cl, K and Mg in the longitudinal walls of columella cells along the lower side of the cap suggest that these ions may move down through the columella tissue of horizontally-oriented roots. Ca also accumulates (largely in the mucilage) along the lower side of the elongating zone of horizontally-oriented roots, while Na, P, Cl and K tend to accumulate along the upper side of the elongating zone. Of these ions, only K increases in concentration in the cytoplasm and longitudinal walls of cortical cells in the upper vs lower sides of the elongating zone. These results indicate that (1) gravity-induced asymmetries of ions differ significantly in the cap and elongating zone of graviresponding roots, (2) Ca accumulates along the lower side of the cap and elongating zone of graviresponding roots, (3) increased growth of the upper side of the elongating zone of horizontally-oriented roots correlates positively with increased amounts of K in the cytoplasm and longitudinal walls of cortical cells, and (4) the apoplast (especially the mucilage) may be an important component of the pathway via which ions move in graviresponding rots of Zea mays. These results are discussed relative to mechanisms for graviperception and gravicurvature of roots.

NASA Program Space Biology↗

Collection of gravitropic effectors from mucilage of electrotropically-stimulated roots of Zea mays L

We placed agar blocks adjacent to tips of electrotropically stimulated primary roots of Zea mays. Blocks placed adjacent to the anode-side of the roots for 3 h induced significant curvature when subsequently placed asymmetrically on tips of vertically-oriented roots. Curvature was always toward the side of the root unto which the agar block was placed. Agar blocks not contacting roots and blocks placed adjacent to the cathode-side of electrotropically stimulated roots did not induce significant curvature when placed asymmetrically on tips of vertically-oriented roots. Atomic absorption spectrophotometry indicated that blocks adjacent to the anode-side of electrotropically-stimulated roots contained significantly more calcium than (1) blocks not contacting roots, and (2) blocks contacting the cathode-side of roots. These results demonstrate the presence of a gradient of endogenous Ca in mucilage of electrotropically-stimulated roots (i.e. roots undergoing gravitropic-like curvature).

NASA Discipline Number 40-10↗

On the nature and origin of the calcium asymmetry arising during gravitropic response in etiolated pea epicotyls

Seven day old etiolated pea epicotyls were loaded symmetrically with 3H-indole 3-acetic acid (IAA) or 45Ca2+, then subjected to 1.5 hours of 1g gravistimulation. Epidermal peels taken from top and bottom surfaces after 90 minutes showed an increase in IAA on the lower side and of Ca2+ on the upper side. Inhibitors of IAA movement (TIBA, 9-hydroxyfluorene carboxylic acid) block the development of both IAA and Ca2+ asymmetries, but substances known to interfere with normal Ca2+ transport (nitrendipine, nisoldipine, Bay K 8644, A 23187) do not significantly alter either IAA or Ca2+ asymmetries. These substances, however, are active in modifying both Ca2+ uptake and efflux through oat and pea leaf protoplast membranes. We conclude that the 45Ca2+ fed to pea epicotyls occurs largely in the cell wall, and that auxin movement is primary and Ca2+ movement secondary in gravitropism. We hypothesize that apoplastic Ca2+ changes during graviresponse because it is displaced by H+ secreted through auxin-induced proton release. This proposed mechanism is supported by localized pH experiments, in which filter paper soaked in various buffers was applied to one side of a carborundum-abraded epicotyls. Buffer at pH 3 increases calcium loss from the side to which it is applied, whereas pH 7 buffer decreases it. Moreover, 10 micromolar IAA and 1 micromolar fusicoccin, which promote H+ efflux, increase Ca2+ release from pea epicotyl segments, whereas cycloheximide, which inhibits H+ efflux, has the reverse effect. We suggest that Ca2+ does not redistribute actively during gravitropism: the asymmetry arises because of its release from the wall adjacent to the region of high IAA concentration, proton secretion, and growth. Thus, the asymmetric distribution of Ca2+ appears to be a consequence of growth stimulation, not a critical step in the early phase of the graviresponse.

NASA Discipline Number 40-10↗

Polar transport of 45Ca2+ across the elongation zone of gravistimulated roots

The movement of calcium across the elongation zone of gravistimulated primary roots of maize (Zea mays L.) was measured using 45Ca2+. Radioactive calcium was applied to one side of the elongation zone about 4 mm back from the root tip and the distribution of radioactivity across the root in the region of application was determined using scintillation spectrometry. The movement of 45Ca2+ across the elongation zone was non-polar in vertically oriented roots. In gravistimulated roots the movement of label was polarized with about twice as much label moving from top to bottom as from bottom to top. A variety of treatments which interfere with gravitropism was found to eliminate the polar movement of 45Ca2+ across the elongation zone. In maize cultivars which require light for gravitropic competency, dark grown roots exhibited neither gravitropism nor polar movement of 45Ca2+ across the elongation zone. Upon illumination the roots developed but gravitropic competency and gravity-induced polar movement of 45Ca2+ across the elongation zone. Similarly, roots of light-grown seedlings lost both gravitropic competency and 45Ca2+ transport polarity upon transfer to the dark. The results indicate a close correlation between calcium movement and gravitropism in primary roots in maize.

NASA Discipline Number 40-10↗

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↗

An electric current associated with gravity sensing in maize roots

The study of gravisensing would be greatly enhanced if physiological events associated with gravity sensing could be detected separately from subsequent growth processes. This report presents a means to discriminate sensing from the growth processes. By using a vibrating probe, we have found an electric current generated by the gravity sensing region of the root cap of maize (Zea mays cv Merit) in response to gravistimulation. On the upper surface of the root cap, the change from the endogenous current has a density of 0.55 microampere per square centimeter away from gravity. The onset of the current shift has a characteristic of lag of three to four minutes after gravistimulation, which corresponds to the presentation time for gravity sensing in this tissue. A description of the current provides some information about the sensing mechanism, as well as being a valuable means to detect gravity sensing independently of differential growth.

NASA Discipline Number 40-10↗

Kinetics of shoot inversion-induced ethylene production in Pharbitis nil

Shoot inversion promotes a significant increase in ethylene production in the inverted part of the Pharbitis nil main shoot. The latent period for shoot inversion-induced ethylene production is ca. 2.75 h. Our results indicate that the shoot-inversion ethylene response is not persistent and can be terminated and rapidly reinitiated by appropriate alteration of the orientation of the main shoot regardless of prolonged previous exposures of the shoot to various orientations. The time course of the production of ACC (1-aminocyclopropane-1-carboxylic acid), the immediate precursor of ethylene, follows a pattern similar to that of ethylene during the various alterations of shoot orientation. Excised stem segments and intact stems are capable of induction, inhibition, and reinduction of ethylene evolution. Ethylene production reported here for shoot inversion does not result from segmenting (wounding) of the tissue.

NASA Discipline Number 40-10↗

Gibberellin-enhanced elongation of inverted Pharbitis nil shoot prevents the release of apical dominance

Ethylene evolution resulting from the gravity stress of shoot inversion appears to induce the release of apical dominance in Pharbitis nil (L.) by inhibiting elongation of the inverted shoot. It has been previously demonstrated that this shoot inversion release of apical dominance can be prevented by promoting elongation in the inverted shoot via interference with ethylene synthesis or action. In the present study it was shown that apical dominance release can also be prevented by promoting elongation of the inverted shoot via treatment with gibberellic acid (GA3). A synergistic effect was observed when AgNO3, the ethylene action inhibitor, was applied with GA3. Both GA3 and AgNO3 increased ethylene production in the inverted shoot. These results are consistent with the view that it is ethylene-induced inhibition of elongation and not any direct effect of ethylene per se which is responsible for the outgrowth of the highest lateral bud.

NASA Discipline Number 40-10↗

Ultrastructural analysis of cell component distribution in the apical cell of Ceratodon protonemata

A distinctive feature of tip-growing plant cells is that cell components are distributed differentially along the length of the cell, although most ultrastructural analyses have been qualitative. The longtitudinal distribution of cell components was studied both qualitatively and quantitatively in the apical cell of dark-grown protonemata of the moss Ceratodon. The first 35 micrometers of the apical cell was analyzed stereologically using transmission electron microscopy. There were four types of distributions along the cell's axis, three of them differential: (1) tubular endoplasmic reticulum was evenly distributed, (2) cisternal endoplasmic reticulum and Golgi vesicles were distributed in a tip-to-base gradient, (3) plastids, vacuoles, and Golgi stacks were enriched in specific areas, although the locations of the enrichments varied, and (4) mitochondria were excluded in the tip-most 5 micrometers and evenly distributed throughout the remaining 30 micrometers. This study provides one of the most comprehensive quantitative, ultrastructural analyses of the distribution of cell components in the apex of any tip-growing plant cell. The finding that almost every component had its own spatial arrangement demonstrates the complexity of the organization and regulation of the distribution of components in tip-growing cells.

Non-NASA Center↗

Gravity related features of plant growth behavior studied with rotating machines

Research in plant physiology consists mostly of studies on plant growth because almost everything a plant does is done by growing. Most aspects of plant growth are strongly influenced by the earth's gravity vector. Research on those phenomena address scientific questions specifically about how plants use gravity to guide their growth processes.

NASA Discipline Plant Biology↗

The response to gravity is correlated with the number of statoliths in Chara rhizoids

In contrast to higher plants, Chara rhizoids have single membrane-bound compartments that appear to function as statoliths. Rhizoids were generated by germinating zygotes of Chara in either soil water (SW) medium or artificial pond water (APW) medium. Differential-interference-contrast microscopy demonstrated that rhizoids form SW-grown plants typically contain 50 to 60 statoliths per cell, whereas rhizoids from APW-grown plants contain 5 to 10 statoliths per cell. Rhizoids from SW are more responsive to gravity than rhizoids from APW because (a) SW rhizoids were oriented to gravity during vertical growth, whereas APW rhizoids were relatively disoriented, and (b) curvature of SW rhizoids was 3 to 4 times greater throughout the time course of curvature. The growth rate of APW rhizoids was significantly greater than that of SW-grown rhizoids. This latter result suggests that APW rhizoids are not limited in their ability for gravitropic curvature by growth and that these rhizoids are impaired in the early stages of gravitropism (i.e. gravity perception). Plants grown in APW appeared to be healthy because of their growth rate and the vigorous cytoplasmic streaming observed in the rhizoids. This study is comparable to earlier studies of gravitropism in starch-deficient mutants of higher plants and provides support for the role of statoliths in gravity perception.

Non-NASA Center↗

The influence of gravity on the formation of amyloplasts in columella cells of Zea mays L

Columella (i.e., putative graviperceptive) cells of Zea mays seedlings grown in the microgravity of outer space allocate significantly less volume to putative statoliths (amyloplasts) than do columella cells of Earth-grown seedlings. Amyloplasts of flight-grown seedlings are significantly smaller than those of ground controls, as is the average volume of individual starch grains. Similarly, the relative volume of starch in amyloplasts in columella cells of flight-grown seedlings is significantly less than that of Earth-grown seedlings. Microgravity does not significantly alter the volume of columella cells, the average number of amyloplasts per columella cell, or the number of starch grains per amyloplast. These results are discussed relative to the influence of gravity on cellular and organellar structure.

NASA Discipline Number 40-10↗

How cereal grass shoots perceive and respond to gravity

The leaf-sheath pulvinus of grasses presents a unique system for studying gravitropism, primarily because of its differences from other organs. The mature pulvinus is a discrete organ specialized for gravitropism: it is nongrowing in the absence of gravistimulation and capable of displaying a graviresponse independent of the rest of the plant. In this paper we present a model for gravitropism in pulvini based on recent findings from studies on the mechanisms of graviperception and graviresponse. According to this model, amyloplasts play an essential role in perceiving a change in the orientation of the pulvinus. The perception of this reorientation leads to the enhanced synthesis and release from conjugate of the auxin IAA, and the increased conjugation of gibberellin, on a localized basis. Because there is a graded growth promotion across the gravistimulated pulvinus, it is suggested that the observed hormonal asymmetry is actually an indication of a linear gradient of hormone concentration, as well as hormone response, across the pulvinus. It is further suggested that the linear gradient of hormone concentration may be predominantly the result of local changes in hormone level, rather than a product of hormonal movement into or across the pulvinus.

NASA Discipline Number 40-99↗