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At least 19 recordsLinked to original sources

Organization of photosystem I polypeptides examined by chemical cross-linking.

Photosystem I from the cyanobacterium Synechocystis sp. PCC 6803 was examined using the chemical cross-linkers glutaraldehyde and N-ethyl-1-3-[3-(dimethylamino)propyl]carbodiimide to investigate the organization of the polypeptide subunits. Thylakoid membranes and photosystem I, which was isolated by Triton X-100 fractionation, were treated with cross-linking reagents and were resolved using a Tricine/urea low-molecular-weight resolution gel system. Subunit-specific antibodies and western blotting analysis were used to identify the components of cross-linked species. These analyses identified glutaraldehyde-dependent cross-linking products composed of small amounts of PsaD and PsaC, PsaC and PsaE, and PsaE and PsaF. The novel cross-link between PsaE and PsaF was also observed following treatment with N-ethyl-1-3-[3-(dimethylamino)propyl]carbodiimide. These cross-linking results suggest a structural interaction between PsaE and PsaF and predict a transmembrane topology for PsaF.

Cross-Linking Reagents↗

Microgravity and clinorotation cause redistribution of free calcium in sweet clover columella cells.

In higher plants, calcium redistribution is believed to be crucial for the root to respond to a change in the direction of the gravity vector. To test the effects of clinorotation and microgravity on calcium localization in higher plant roots, sweet clover (Melilotus alba L.) seedlings were germinated and grown for two days on a slow rotating clinostat or in microgravity on the US Space Shuttle flight STS-60. Subsequently, the tissue was treated with a fixative containing antimonate (a calcium precipitating agent) during clinorotation or in microgravity and processed for electron microscopy. In root columella cells of clinorotated plants, antimonate precipitates were localized adjacent to the cell wall in a unilateral manner. Columella cells exposed to microgravity were characterized by precipitates mostly located adjacent to the proximal and lateral cell wall. In all treatments some punctate precipitates were associated with vacuoles, amyloplasts, mitochondria, and euchromatin of the nucleus. A quantitative study revealed a decreased number of precipitates associated with the nucleus and the amyloplasts in columella cells exposed to microgravity as compared to ground controls. These data suggest that roots perceive a change in the gravitational field, as produced by clinorotation or space flights, and respond respectively differently by a redistribution of free calcium.

Antimony↗

Up-regulation of phosphoinositide metabolism in tobacco cells constitutively expressing the human type I inositol polyphosphate 5-phosphatase.

To evaluate the impact of suppressing inositol 1,4,5-trisphosphate (InsP(3)) in plants, tobacco (Nicotiana tabacum) cells were transformed with the human type I inositol polyphosphate 5-phosphatase (InsP 5-ptase), an enzyme which specifically hydrolyzes InsP(3). The transgenic cell lines showed a 12- to 25-fold increase in InsP 5-ptase activity in vitro and a 60% to 80% reduction in basal InsP(3) compared with wild-type cells. Stimulation with Mas-7, a synthetic analog of the wasp venom peptide mastoparan, resulted in an approximately 2-fold increase in InsP(3) in both wild-type and transgenic cells. However, even with stimulation, InsP(3) levels in the transgenic cells did not reach wild-type basal values, suggesting that InsP(3) signaling is compromised. Analysis of whole-cell lipids indicated that phosphatidylinositol 4,5-bisphosphate (PtdInsP(2)), the lipid precursor of InsP(3), was greatly reduced in the transgenic cells. In vitro assays of enzymes involved in PtdInsP(2) metabolism showed that the activity of the PtdInsP(2)-hydrolyzing enzyme phospholipase C was not significantly altered in the transgenic cells. In contrast, the activity of the plasma membrane PtdInsP 5 kinase was increased by approximately 3-fold in the transgenic cells. In vivo labeling studies revealed a greater incorporation of (32)P into PtdInsP(2) in the transgenic cells compared with the wild type, indicating that the rate of PtdInsP(2) synthesis was increased. These studies show that the constitutive expression of the human type I InsP 5-ptase in tobacco cells leads to an up-regulation of the phosphoinositide pathway and highlight the importance of PtdInsP(2) synthesis as a regulatory step in this system.

Cells, Cultured↗

Effects of clinorotation and microgravity on sweet clover columella cells treated with cytochalasin D.

The cytoskeleton of columella cells is believed to be involved in maintaining the developmental polarity of cells observed as a reproducible positioning of cellular organelles. It is also implicated in the transduction of gravitropic signals. Roots of sweet clover (Melilotus alba L.) seedlings were treated with a microfilament disrupter, cytochalasin D, on a slowly rotating horizontal clinostat (2 rpm). Electron micrographs of treated columella cells revealed several ultrastructural effects including repositioning of the nucleus and the amyloplasts and the formation of endoplasmic reticulum (ER) whorls. However, experiments performed during fast clinorotation (55 rpm) showed an accumulation (but no whorling) of a disorganized ER network at the proximal and distal pole and a random distribution of the amyloplasts. Therefore, formation of whorls depends upon the speed of clinorotation, and the overall impact of cytochalasin D suggests the necessity of microfilaments in organelle positioning. Interestingly, a similar drug treatment performed in microgravity aboard the US Space Shuttle Endeavour (STS-54, January 1993) caused a displacement of ER membranes and amyloplasts away from the distal plasma membrane. In the present study, we discuss the role of microfilaments in maintaining columella cell polarity and the utility of clinostats to simulate microgravity.

Actin Cytoskeleton↗

Gravitropism of hypocotyls of wild-type and starch-deficient Arabidopsis seedlings in spaceflight studies.

The major purpose of this spaceflight project was to investigate the starch-statolith hypothesis for gravity perception, and a secondary goal was to study plant growth and development under spaceflight conditions. This research was based on our ground studies of gravity perception in the wild type and three starch-deficient (one starchless and two reduced starch) mutants of Arabidopsis thaliana (L.) Heynh. Dark-grown seedlings that developed in microgravity were given one of several (30 min, 60 min, or 90 min) 1-g stimuli by an on-board centrifuge, and additional controls for seedling development also were performed. These latter control experiments included a morphological study of plants that developed in space in microgravity (F microg), in space on a centrifuge (F 1g), on the ground (G 1g), and on a rotating clinostat on the ground. Since elevated levels of ethylene were reported in the spacecraft atmosphere, additional controls for morphology and gravitropism with added ethylene also were performed. While exogenous ethylene reduced the absolute magnitude of the response in all four strains of Arabidopsis, this gas did not appear to change the relative graviresponsiveness among the strains. The relative response of hypocotyls of microgravity-grown seedlings to the stimuli provided by the in-flight centrifuge was: wild type > starch-deficient mutants. Although the protoplast pressure model for gravity perception cannot be excluded, these results are consistent with a statolith-based model for perception in plants.

Arabidopsis↗

Contemplating the plasmalemmal control center model.

An abundant epidermal mechanosensory calcium-selective ion channel appears able not only to detect mechanical stimuli such as those that initiate gravitropism but also to detect thermal, electrical, and various chemical stimuli. Because it responds to multimodal input with a second messenger output, this channel system seems likely to be an integrator that can engage in feedbacks with many other systems of the cell--and feedback is the hallmark of regulation. In general, the mechanical tension required for channel activation is likely transmitted from the relatively rigid cell wall to the plasma membrane system via linkage or adhesion sites that display antigenicities recognized by antibodies to animal beta-1 integrin, vitronectin, and fibronectin and which have mechanical connections to the cytoskeleton. Thus, functionally, leverage exerted against any given adhesion site will tend to control channels within a surrounding domain. Reactions initiated by passage of calcium ions through the channels could presumably be more effectively regulated if channels within the domains were somewhat clustered and if appropriate receptors, kinases, porters, pumps, and some key cytoskeletal anchoring sites were in turn clustered about them. Accumulating evidence suggests not only that activity of clusters of channels may contribute to control of cytoskeletal architecture and of regulatory protein function within their domain, but also that both a variety of regulatory proteins and components of the cortical cytoskeleton may contribute to control of channel activity. The emerging capabilities of electronic optical microscopy are well suited for resolving the spatial distributions of many of these cytoskeletal and regulatory molecules in living cells, and for following some of their behaviors as channels are stimulated to open and cytosolic calcium builds in their vicinity. Such microscopy, coupled with biochemical and physiological probing, should help to establish the nature of the feedback loops putatively controlled by the linkage sites and their channel domains.

Calcium Channels↗

Analysis of multiple photoreceptor pigments for phototropism in a mutant of Arabidopsis thaliana.

The shape of the fluence-response relationship for the phototropic response of the JK224 strain of Arabidopsis thaliana depends on the fluence rate and wavelength of the actinic light. At low fluence rate (0.1 micromole m-2 s-1), the response to 450-nm light is characterized by a single maximum at about 9 micromoles m-2. At higher fluence rate (0.4 micromole m-2 s-1), the response shows two maxima, at 4.5 and 9 micromoles m-2. The response to 510-nm light shows a single maximum at 4.5 micromoles m-2. Unilateral preirradiation with high fluence rate (25 micromoles m-2 s-1) 510-nm light eliminates the maximum at 4.5 micromoles m-2 in the fluence response curve to a subsequent unilateral 450-nm irradiation, while the second maximum at 9 micromoles m-2 is unaffected. Based on these results, it is concluded that a single photoreceptor pigment has been altered in the JK224 strain of Arabidopsis thaliana.

Arabidopsis↗

Some characteristics of photosynthetic apparatus under conditions of spaceflight.

During colonization of space by humans, the bioregenerative life-support systems on board the space ships will require the plants with a highly efficient photosynthesis, a process producing food and O2 and removing CO2-Therefore, in recent years the scientists increasingly focus the their attention to study on photosynthetic apparatus of plants grown in space. Although the available data are quite scanty and, at times, controversial, it is Considered that the space grown plants differ from around control plants by growth and development, many structural and functional indices and metabolism. Data exist showing changes in the chlorophyll (Chl) content, structure and number of chloroplasts in the cell, swelling of thylakoids and decrease in the number and size of starch grains in the chloroplasts (for reviews, see Halstead and Dutcher, 1987; Kordyum, 1997). The decrease of shoot fresh weight and reduction of CO2-saturated photosynthetic rate at saturating light intensities in space grown wheat plants in comparison with ground control have been reported by Tripathy et al. (1996). The thylakoids isolated from space grown plants displayed lower rates of electron transport through photosystems I and II (PSI and PSII) and in a whole chain. This study aimed to examine the electron transport rates through PSI and PSII in the isolated thylakoids, to elucidate if there are any differences in accumulation of thylakoid membranes between space grown Brassica rapa plants and ground control plants (based on Chl quantity) and to measure the Chl a/b ratio in isolated thylakoids. These studies were part of the Collaborative US/Ukrainian program during the STS-87 mission (1997).

Brassica↗

[The evaluation of the error of the thermal pulse technique used to measure moisture content of root substrates in space greenhouse].

Point thermopulse probes were used to monitor moisture level in the root substrates during cultivation of higher plants in a space greenhouse. Investigated were performance data of the thermopulse moisture probe in integration with the space greenhouse. It was shown that within the substrate moisture range from 20 up to 100% of the full saturation the technique error does not exceed 1.5%. The thermopulse technique bears much promise for metrologic monitoring of the root substrate moisture content in space greenhouses no matter water and air supply technology.

Differential Thermal Analysis↗

Mechanism of rapid suppression of cell expansion in cucumber hypocotyls after blue-light irradiation.

Rapid suppression of hypocotyl elongation by blue light in cucumber (Cucumis sativus L.) was studied to examine possible hydraulic and wall changes responsible for diminished growth. Cell-sap osmotic pressure, measured by vapor-pressure osmometry, was not decreased by blue light; turgor pressure, measured by the pressure-probe technique, remained constant during the growth inhibition; and stem hydraulic conductance, measured by dynamic and static methods, was likewise unaffected by blue light. Wall yielding properties were assessed by the pressure-block technique for in-vivo stress relaxation. Blue light reduced the initial rate of relaxation by 77%, but had little effect on the final amount of relaxation. The results demonstrate that blue irradiation acts to decrease the wall yielding coefficient, but not the yield threshold. Stress-strain (Instron) analysis showed that irradiation of the seedlings had little effect on the mechanical extensibilities of the isolated wall. The results indicate that blue light can reduce cell-wall loosening without affecting bulk viscoelastic properties, and indicate a chemorheological mechanism of cell-wall expansion.

Biophysical Phenomena↗

The possible involvement of root-cap mucilage in gravitropism and calcium movement across root tips of Allium cepa L.

Roots of Allium cepa L. grown in aerated water elongate rapidly, but are not graviresponsive. These roots (1) possess extensive columella tissues comprised of cells containing numerous sedimented amyloplasts, (2) lack mucilage on their tips, and (3) are characterized by a weakly polar movement of calcium (Ca) across their tips. Placing roots in humid air correlates positively with the (1) onset of gravicurvature, (2) appearance of mucilage on tips of the roots, and (3) onset of the ability to transport Ca polarly to the lower side of the root tip. Gravicurvature of roots previously submerged in aerated water is more rapid when roots are oriented vertically for 1-2 h in humid air prior to being oriented horizontally. The more rapid gravicurvature of these roots correlates positively with the accumulation of mucilage at the tips of roots during the time the roots are oriented vertically. Therefore, the onset of gravicurvature and the ability of roots to transport Ca to the lower sides of their tips correlate positively with the presence of mucilage at their tips. These results suggest that mucilage may be important for the transport of Ca across root caps.

Allium↗

Graviresponsiveness of surgically altered primary roots of Zea mays.

We examined the gravitropic responses of surgically altered primary roots of Zea mays to determine the route by which gravitropic inhibitors move from the root tip to the elongating zone. Horizontally oriented roots, from which a 1-mm-wide girdle of epidermis plus 2-10 layers of cortex were removed from the apex of the elongating zone, curve downward. However, curvature occurred only apical to the girdle. Filling the girdle with mucilage-like material transmits curvature beyond the girdle. Vertically oriented roots with a half-girdle' (i.e. the epidermis and 2-10 layers of the cortex removed from half of the circumference of the apex of the elongating zone) curve away from the girdle. Inserting the half-girdle at the base of the elongating zone induces curvature towards the girdle. Filling the half-circumference girdles with mucilage-like material reduced curvature significantly. Stripping the epidermis and outer 2-5 layers of cortex from the terminal 1.5 cm of one side of a primary root induces curvature towards the cut, irrespective of the root's orientation to gravity. This effect is not due to desiccation since treated roots submerged in water also curved towards their cut surface. Coating a root's cut surface with a mucilage-like substance minimizes curvature. These results suggest that the outer cell-layers of the root, especially the epidermis, play an important role in root gravicurvature, and the gravitropic signals emanating from the root tip can move apoplastically through mucilage.

Biological Transport↗

Wall relaxation and the driving forces for cell expansive growth.

When water uptake by growing cells is prevented, the turgor pressure and the tensile stress in the cell wall are reduced by continued wall loosening. This process, termed in vivo stress relaxation, provides a new way to study the dynamics of wall loosening and to measure the wall yield threshold and the physiological wall extensibility. Stress relaxation experiments indicate that wall stress supplies the mechanical driving force for wall yielding. Cell expansion also requires water absorption. The driving force for water uptake during growth is created by wall relaxation, which lowers the water potential of the expanding cells. New techniques for measuring this driving force show that it is smaller than believed previously; in elongating stems it is only 0.3 to 0.5 bar. This means that the hydraulic resistance of the water transport pathway is small and that rate of cell expansion is controlled primarily by wall loosening and yielding.

Biological Transport↗

The relationship between water binding and desiccation tolerance in tissues.

In an effort to define the nature of desiccation tolerance, a comparison of the water sorption characteristics was made between tissues that were resistant and tissues that were sensitive to desiccation. Water sorption isotherms were constructed for germinated and ungerminated soybean axes and also for fronds of several species of Polypodium with varying tolerance to dehydration. The strength of water binding was determined by van't Hoff as well as D'Arcy/Watt analyses of the isotherms at 5, 15, and/or 25 degrees C. Tissues which were sensitive to desiccation had a poor capacity to bind water tightly. Tightly bound water can be removed from soybean and pea seeds by equilibration at 35 degrees C over very low relative humidities; this results in a reduction in the viability of the seed. We suggest that region 1 water (i.e. water bound with very negative enthalpy values) is an important component of desiccation tolerance.

Desiccation↗

Tissue to tissue symplastic communication in the shoots of etiolated corn seedlings.

Carboxyfluorescein, a symplastic probe, was applied to the cut mesocotyl base or coleoptile apex of etiolated Zea mays cv. Silver Queen seedlings and its transport measured and tissue distribution determined. Long-distance longitudinal symplastic transport of the carboxyfluorescein was mainly in the vascular stele. It moved laterally from the mesocotyl stele to the mesocotyl cortex but the presence of a weak barrier limited the movement. A partial symplastic barrier was also present near the coleoptile-mesocotyl node.

Biological Transport↗

A gradient of endogenous calcium forms in mucilage of graviresponding roots of Zea mays.

Agar blocks that contacted the upper sides of tips of horizontally-oriented roots of Zea mays contain significantly less calcium (Ca) than blocks that contacted the lower sides of such roots. This gravity-induced gradient of Ca forms prior to the onset of gravicurvature, and does not form across tips of vertically-oriented roots or roots of agravitropic mutants. These results indicate that (1) Ca can be collected from mucilage of graviresponding roots, (2) gravity induces a downward movement of endogenous Ca in mucilage overlying the root tip, (3) this gravity-induced gradient of Ca does not form across tips of agravitropic roots, and (4) formation of a Ca gradient is not a consequence of gravicurvature. These results are consistent with gravity-induced movement of Ca being a trigger for subsequent redistribution of growth effectors (e.g. auxin) that induce differential growth and gravicurvature.

Biological Transport↗