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At least 37 records · Page 2

Diversity and regulation of plant Ca2+ pumps: insights from expression in yeast

The spatial and temporal regulation of calcium concentration in plant cells depends on the coordinate activities of channels and active transporters located on different organelles and membranes. Several Ca2+ pumps have been identified and characterized by functional expression of plant genes in a yeast mutant (K616). This expression system has opened the way to a genetic and biochemical characterization of the regulatory and catalytic features of diverse Ca2+ pumps. Plant Ca(2+)-ATPases fall into two major types: AtECA1 represents one of four or more members of the type IIA (ER-type) Ca(2+)-ATPases in Arabidopsis, and AtACA2 is one of seven or more members of the type IIB (PM-type) Ca(2+)-ATPases that are regulated by a novel amino terminal domain. Type IIB pumps are widely distributed on membranes, including the PM (plasma membrane), vacuole, and ER (endoplasmic reticulum). The regulatory domain serves multiple functions, including autoinhibition, calmodulin binding, and sites for modification by phosphorylation. This domain, however, is considerably diverse among several type IIB ATPases, suggesting that the pumps are differentially regulated. Understanding of Ca2+ transporters at the molecular level is providing insights into their roles in signaling networks and in regulating fundamental processes of cell biology.

Review, Academic↗

Strontium transfer from maternal skeleton to the fetus estimated on the basis of the Techa river data

Measurements of 90Sr in human bone of inhabitants of the Techa river region were started in 1951, and since 1974 the Techa river population has been studied with a whole-body counter. One of the dosimetric tasks that could be decided using data on 90Sr measurements is direct evaluation of strontium transfer to the fetus from the maternal skeleton. Six cases were selected for which 90Sr measurements were available both for stillborn infants and their mothers. The ratio of 90Sr concentrations in fetal bone to maternal bone for the year of pregnancy has been evaluated. Two clusters of values were found and the difference between clusters could be explained by age-dependent features of maternal bone formation and remodelling. When the mother's 90Sr intake occurred in the period of intensive compact bone growth, the transfer coefficient was very low (0.012-0.032). If 90Sr ingestion occurred during the woman's reproductive age, the transfer to fetus was equal to 0.21-0.26.

Non-NASA Center↗

Optimizing the calcium content of a copolymer acrylamide gel matrix for dark-grown seedlings

A copolymer acrylamide acrylate gel was investigated as the sole root matrix for dark-grown seedlings of soybean (Glycine max Merr. 'Century 84'). Increasing Ca2+ in the hydrating solution of the hydrogel from 1 to 10 mM decreased its water-holding capacity from 97 to 46 mL g-1, yet water potential of the medium remained high, sufficient for normal plant growth at all Ca2+ concentrations tested. Elongation rate of dark-grown soybean seedlings over a 54-hour period was 0.9, 1.5, and 1.8 mm h-1 with 1.0, 2.5, or 5.0 mM Ca2+, respectively, but did not increase with further increases in Ca2+ concentration. Further study revealed that Na+ was released from the hydrogel medium and was taken up by the seedlings as Ca2+ increased in the medium. In dry hypocotyl tissue, sodium content correlated negatively with calcium content. Despite the presence of Na+ in the hydrogel, seedling growth was normal when adequate Ca2+ was added in the hydrating solution. Acrylamide hydrogels hold good potential as a sole growth matrix for short-term experiments with dark-grown seedlings without irrigation.

NASA Discipline Plant Biology↗

The endomembrane sheath: a key structure for understanding the plant cell?

Recent evidence suggests that integrin is abundant in endomembranes of plant cells, and the endomembranes are clad by a sheath of cytoskeleton including F-actin. A role for endomembrane integrin and the endomembrane sheath is proposed: this system might orchestrate metabolic regulation by providing and modulating loci for channelling, and might accelerate channeling as needed by dragging the endoplasmic reticulum (ER) and organelles through the cytoplasm. To accomplish this "streaming", F-actin might lever against the rest of the endomembrane sheath and the ER might also lever against adhesion sites (i.e., plasmodesmata and plasmalemmal control centers). As an important agent in the control of cellular activities, according to this model, the endomembrane sheath would play a major part in responses to diverse signals and stresses, and under extreme stress cell survival would depend on the ability of the system to maintain enough integrity to direct critical syntheses and degradations.

NASA Discipline Plant Biology↗

Recycling crop residues for use in recirculating hydroponic crop production

As part of bioregenerative life support feasibility testing by NASA, crop residues are being used to resupply elemental nutrients to recirculating hydroponic crop production systems. Methods for recovering nutrients from crop residues have evolved from water soaking (leaching) to rapid aerobic bioreactor processing. Leaching residues recovered the majority of elements but it also recovered significant amounts of soluble organics. The high organic content of leachates was detrimental to plant growth. Aerobic bioreactor processing reduced the organic content ten-fold, which reduced or eliminated phytotoxic effects. Wheat and potato production studies were successful using effluents from reactors having with 8- to 1-day retention times. Aerobic bioreactor effluents supplied at least half of the crops elemental mass needs in these studies. Descriptions of leachate and effluent mineral content, biomass productivity, microbial activity, and nutrient budgets for potato and wheat are presented.

NASA Center KSC↗

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

The distribution of calcium (Ca) in caps of vertically- and horizontally-oriented roots of Zea mays was monitored to determine its possible role in root graviresponsiveness. A modification of the antimonate precipitation procedure was used to localize Ca in situ. In vertically-oriented roots, the presumed graviperceptive (i.e., columella) cells were characterized by minimal and symmetric staining of the plasmalemma and mitochondria. No precipitate was present in plasmodesmata or cell walls. Within 5 min after horizontal reorientation, staining was associated with the portion of the cell wall adjacent to the distal end of the cell. This asymmetric staining persisted throughout the onset of gravicurvature. No staining of lateral cell walls of columella cells was observed at any stage of gravicurvature, suggesting that a lateral flow of Ca through the columella tissue of horizontally-oriented roots does not occur. The outermost peripheral cells of roots oriented horizontally and vertically secrete Ca through plasmodesmata-like structures in their cell walls. These results are discussed relative to proposed roles of root-cap Ca in root gravicurvature.

NASA Discipline Plant Biology↗

Nutrition in space

The authors review studies conducted to define nutritional requirements for astronauts during space flight and to assess nutrition before, during, and after space flight. Topics include space food systems, research and limitations on spacecraft, physiological adaptation to weightlessness, energy requirements, dietary intake during space flight, bone demineralization, gastrointestinal function, blood volume, and nutrition requirements for space flight. Benefits of space-related nutrition research are highlighted.

Review↗

Molecular genetics of root gravitropism and waving in Arabidopsis thaliana

When Arabidopsis thaliana seedlings grow embedded in an agar-based medium, their roots grow vertically downward. This reflects their ability to sense the gravity vector and to position their tip parallel to it (gravitropism). We have isolated a number of mutations affecting root gravitropism in Arabidopsis thaliana. One of these mutations, named arg1, affects root and hypocotyl gravitropism without promoting defects in starch content or in the ability of seedlings' organs to respond to plant hormones. The ARG1 gene was cloned and shown to code for a protein with a J domain at its amino terminus and a second sequence motif found in several cytoskeleton binding proteins. Mutations in the AGR1 locus promote a strong defect in root gravitropism. Some alleles also confer an increased root resistance to exogenous ethylene and an increased sensitivity to auxin. AGR1 was cloned and found to encode a putative transmembrane protein which might be involved in polar auxin transport, or in regulating the differential growth response to gravistimulation. When Arabidopsis seedlings grow on the surface of agar-based media tilted backward, their roots wave. That wavy pattern of root growth derives from a combined response to gravity, touch and other surface-derived stimuli. It is accompanied by a reversible rotation of the root tip about its axis. A number of mutations affect the presence or the shape of root waves on tilted agar-based surfaces. One of them, wvc1, promotes the formation of compressed root waves under these conditions. The physiological and molecular analyses of this mutant suggest that a tryptophan-derived molecule other than IAA might be an important regulator of the curvature responsible for root waving.

NASA Discipline Plant Biology↗

Effects of artificial enclosure of young lettuce leaves on tipburn incidence and leaf calcium concentration

The young developing leaves of 20-day-old lettuce plants (Lactuca sativa L. 'Buttercrunch') were enclosed by aluminized polyethylene sheaths to decrease transpiration and reduce Ca transport. The plants were grown in recirculating solution culture system using a modified half-strength Hoagland's solution under cool-white fluorescent lamps with a photosynthetic photon flux of 350 micromoles s-1 m-2 in a 16:8-hr (light:dark) period. Air temperature and humidity were 20 degrees C and 65%, respectively. After 4 days of enclosure, 53% of the inner leavers (leaves one to 3 cm in length) were tipburned. After the same period, less than 1% of the inner leaves on control plants were tipburned. The concentration of Ca in enclosed inner leaves was 0.63 mg g-1 dry weight, compared to 1.48 mg g-1 dry weight in inner leaves that were not enclosed. The Ca concentration in transpiring outer leaves of all plants was 9.9 mg g-1 dry weight. The Mg concentration in enclosed inner leaves was 2.25 mg g-1 dry weight, compared to 2.34 mg g-1 dry weight in inner leaves that were not enclosed. This research documents that enclosures of leaves at the growing point, as would occur with normal head development, is sufficient to create a limiting concentration of Ca in the enclosed tissue and encourage tipburn development.

Non-NASA Center↗

Statoconia formation in molluscan statocysts

The gravity sensors of all molluscs phylogenetically below the cephalopods are spherical organs called statocysts. The wall of the sphere contains mechanosensory cells whose sensory cilia project into the lumen of the cyst. The lumen is filled with fluid and dense "stones", the statoconia or statoliths, which sink under the influence of gravity to load, and stimulate, those receptor cells which are at the bottom. The statoconia of Aplysia californica are shown to be calcified about a lamellar arrangement of membranes. Similar lamellar membrane arrangements are seen within the receptor cells, and their possible role in the formation of the statoconia is discussed. SEM of unfixed statoconia reveals plate-like crystallization on their surface. Elemental analysis shows a relatively high Sr content, which is of interest, since others have recently reported that Sr is required in the culture medium of several laboratory reared molluscs in order for the statoconia to develop.

Non-NASA Center↗

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↗

Calbindins decreased after space flight

Exposure of the body to microgravity during space flight causes a series of well-documented changes in Ca2+ metabolism, yet the cellular and molecular mechanisms leading to these changes are poorly understood. Calbindins, vitamin D-dependent Ca2+ binding proteins, are believed to have a significant role in maintaining cellular Ca2+ homeostasis. In this study, we used biochemical and immunocytochemical approaches to analyze the expression of calbindin-D28k and calbindin-D9k in kidneys, small intestine, and pancreas of rats flown for 9 d aboard the space shuttle. The effects of microgravity on calbindins in rats from space were compared with synchronous Animal Enclosure Module controls, modeled weightlessness animals (tail suspension), and their controls. Exposure to microgravity resulted in a significant and sustained decrease in calbindin-D28k content in the kidney and calbindin-D9k in the small intestine of flight animals, as measured by enzyme-linked immunosorbent assay (ELISA). Modeled weightlessness animals exhibited a similar decrease in calbindins by ELISA. Immunocytochemistry (ICC) in combination with quantitative computer image analysis was used to measure in situ the expression of calbindins in the kidney and the small intestine, and the expression of insulin in pancreas. There was a large decrease of immunoreactivity in renal distal tubular cell-associated calbindin-D28k and in intestinal absorptive cell-associated calbindin-D9k of space flight and modeled weightlessness animals compared with matched controls. No consistent difference in pancreatic insulin immunoreactivity between space flight, modeled weightlessness, and controls was observed. Regression analysis of results obtained by quantitative ICC and ELISA for space flight, modeled weightlessness animals, and their controls demonstrated a significant correlation. These findings after a short-term exposure to microgravity or modeled weightlessness suggest that a decreased expression of calbindins may contribute to the disorders of Ca2+ metabolism induced by space flight.

Non-NASA Center↗

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.

NASA Discipline Number 40-10↗

Potassium concentration effect on growth, gas exchange and mineral accumulation in potatoes

This study was conducted to evaluate the responses of potatoes to six K solution concentrations maintained with a flow-through nutrient film system. Potato plants were grown for 42 days in sloping shallow trays containing a 1 cm layer of quartz gravel with a continuous flow of 4 ml min-1 of nutrient solutions having K concentrations of 0.10, 0.55, 1.59, 3.16, 6.44, 9.77 meq L-1. Plant leaf area, total and tuber dry weights were reduced over 25% at 0.10 meq L-1 of K and over 17% at 9.77 meq L-1 of K compared to concentrations of 0.55, 1.59, 3.16 and 6.44 meq L-1 of K. Gas exchange measurements on leaflets in situ after 39 days of growth demonstrated no significant differences among different K treatments in CO2 assimilation rate, stomatal conductance, intercellular CO2 concentration, and transpiration. Further measurements made only on plants grown at 0.10, 1.59, 6.44 meq L-1 of K showed similar responses of CO2 assimilation rate to different intercellular CO2 concentrations. This suggested that the photosynthetic systems were not affected by different K nutrition. The leaves of plants accumulated about 60% less K at 0.10 meq L-1 of K than at higher K concentrations. However, Ca and Mg levels in the leaves were higher at 0.10 meq L-1 of K than at higher K concentrations. This indicates that low K nutrition not only reduced plant growth, but also affected nutrient balance between major cations.

Non-NASA Center↗

Rapid changes in protein phosphorylation associated with light-induced gravity perception in corn roots

The effect of light and calcium depletion on in vivo protein phosphorylation was tested using dark-grown roots of Merit corn. Light caused rapid and specific promotion of phosphorylation of three polypeptides. Pretreatment of roots with ethylene glycol bis N,N,N',N' tetraacetic acid and A23187 prevented light-induced changes in protein phosphorylation. We postulate that these changes in protein phosphorylation are involved in the light-induced gravity response.

NASA Program Space Biology↗

Partial purification and characterization of a Ca(2+)-dependent protein kinase from pea nuclei

Almost all the Ca(2+)-dependent protein kinase activity in nuclei purified from etiolated pea (Pisum sativum, L.) plumules is present in a single enzyme that can be extracted from chromatin by 0.3 molar NaCl. This protein kinase can be further purified 80,000-fold by salt fractionation and high performance liquid chromatography, after which it has a high specific activity of about 100 picomoles per minute per microgram in the presence of Ca2+ and reaches half-maximal activation at about 3 x 10(-7) molar free Ca2+, without calmodulin. It is a monomer with a molecular weight near 90,000. It can efficiently use histone III-S, ribosomal S6 protein, and casein as artificial substrates, but it phosphorylates phosvitin only weakly. Its Ca(2+)-dependent kinase activity is half-maximally inhibited by 0.1 millimolar chlorpromazine, by 35 nanomolar K-252a and by 7 nanomolar staurosporine. It is insensitive to sphingosine, an inhibitor of protein kinase C, and to basic polypeptides that block other Ca(2+)-dependent protein kinases. It is not stimulated by exogenous phospholipids or fatty acids. In intact isolated pea nuclei it preferentially phosphorylates several chromatin-associated proteins, with the most phosphorylated protein band being near the same molecular weight (43,000) as a nuclear protein substrate whose phosphorylation has been reported to be stimulated by phytochrome in a calcium-dependent fashion.

NASA Discipline Number 40-50↗

Movement of endogenous calcium in the elongating zone of graviresponding roots of Zea mays

Endogenous calcium (Ca) accumulates along the lower side of the elongating zone of horizontally oriented roots of Zea mays cv. Yellow Dent. This accumulation of Ca correlates positively with the onset of gravicurvature, and occurs in the cytoplasm, cell walls and mucilage of epidermal cells. Corresponding changes in endogenous Ca do not occur in cortical cells of the elongating zone of intact roots. These results indicate that the calcium asymmetries associated with root gravicurvature occur in the outermost layers of the root.

NASA Discipline Number 29-20↗