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| Körner, L E, Kjellbom, P, Larsson, C, Møller, I M | authors |
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| Isolation of Bundle Sheath Cell Chloroplasts from the NADP-ME Type C(4) Plant Zea mays: Capacities for CO(2) Assimilation and Malate Decarboxylation. | name |
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| Bundle sheath chloroplasts have been isolated from Zea mays leaves by a procedure involving enzymic digestion of mechanically prepared strands of bundle sheath cells followed by gentle breakage and filtration. The resulting crude chloroplast preparation was enriched by Percoll density layer centrifugation to yield intact chloroplasts (about 20 micrograms chlorophyll per 10-gram leaf tissue) with high metabolic activities. Based on activities of marker enzymes in the chloroplast and bundle sheath cell extracts, the chloroplasts were essentially free of contamination by other organelles and cytoplasmic material, and were generally about 70% intact. Chlorophyll a/b ratios were high (about 10). With appropriate substrates these chloroplasts displayed high rates of malate decarboxylation, measured as pyruvate formation, and CO(2) assimilation (maximum rates approximately 5 and 3 micromoles per minute per milligram chlorophyll, respectively). These activities were light dependent, linear for at least 20 minutes at 30 degrees C, and displayed highest rates at pH 8.0. High metabolic rates were dependent on addition of an exogenous source of carbon to the photosynthetic carbon reduction cycle (3-phosphoglycerate or dihydroxyacetone phosphate) and a nucleotide (ATP, ADP, or AMP), as well as aspartate. Generally, neither malate decarboxylation nor CO(2) assimilation occurred substantially in the absence of the other activity indicating a close relationship between these processes. Presumably, NADPH required for the photosynthetic carbon reduction cycle is largely supplied during the decarboxylation of malate by NADP-malic enzyme. The results are discussed in relation to the role of bundle sheath chloroplasts in C(4) photosynthesis by species of the NADP-malic enzyme type. | description |
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| Jenkins, C L, Boag, S | authors |
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| Studies on Conditions for Cell Division and Embryogenesis in Isolated Pollen Culture of Nicotiana rustica. | name |
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| A method for the induction of a high rate of cell division and embryogenesis of Nicotiana rustica pollen was developed. Binucleate pollen grains were fractionated by Percoll density gradient (35/45%) centrifugation and cultured in 0.4 molar mannitol at 30 degrees C (the first culture). After 3 days in culture pollen was recollected by a second Percoll fractionation (0/30%) and transferred to and cultured in a medium containing the Murashige-Skoog macro-elements, 0.4 molar mannitol, 40 millimolar galactose, 3 millimolar glutamine, and 5 micromolar ABA for 10 days (the second culture). The cell population consisting of about 80% dividing pollen was transferred to a Murashige-Skoog medium containing 0.4 molar mannitol, 3 millimolar glutamine, and no phytohormone (the third culture), where about 40% of dividing pollen developed into embryos or embryogenic calli. | description |
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| Kyo, M, Harada, H | authors |
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| Isolation of serine:glyoxylate aminotransferase from cucumber cotyledons. | name |
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| Serine:glyoxylate aminotransferase, a marker enzyme for leaf peroxisomes, has been purified to homogeneity from cucumber cotyledons (Cucumis sativus cv Improved Long Green). The isolation procedure involved precipitation with polyethyleneimine, a two-step ammonium sulfate fractionation (35 to 45%), gel filtration on Ultrogel AcA 34, and ion exchange chromatography on diethylaminoethyl-cellulose, first in the presence of pyridoxal-5-phosphate, and then in its absence. The enzyme was purified approximately 690-fold to a final specific activity of 34.4 units per milligram. Electrophoresis of the purified enzyme on sodium dodecyl sulfate-polyacrylamide gels revealed two polypeptide bands with apparent molecular weights of approximately 47,000 and 45,000. Both polypeptides coeluted with enzyme activity under all chromatographic conditions investigated, both were localized to the peroxisome, and both accumulated in cotyledons as enzyme activity increased during development. The two polypeptides appear not to be structurally related, since they showed little immunological cross-reactivity and gave rise to different peptide fragments when subjected to partial proteolytic digestion. Antiserum raised against either the denatured enzyme or the 45,000-dalton polypeptide did not react with any other polypeptides present in a crude cotyledonary homogenate. The purified enzyme also had alanine:glyoxylate aminotransferase activity, but was about twice as active with serine as the amino donor. | description |
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| Hondred, D, Hunter, J M, Keith, R, Titus, D E, Becker, W M | authors |
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| Leaf k interaction with water stress inhibition of nonstomatal-controlled photosynthesis. | name |
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| The relationship between leaf K(+) concentration, in vitro dehydration, and nonstomatal-controlled photosynthesis was investigated using leaf slices that were vacuum infiltrated with media containing varying sorbitol concentrations. The leaf slices were from plants either supplied with complete or K(+)-deficient medium throughout a 35-day growth period. During this time, leaf K(+) concentration, water potential, osmotic potential, and turgor pressure were monitored. Leaf K(+) concentration averaged 239 micomoles per gram (fresh weight) in control plants, and dropped to 74.3 micromoles per gram (fresh weight) in K(+)-deficient plants. Less negative osmotic potentials and resultant turgor loss in K(+)-deficient plants indicated that the osmotically active pool of cellular K(+) was lower in those plants.The decrease in leaf K(+) concentration enhanced the dehydration inhibition of photosynthesis. For example, increasing sorbitol from 0.33 to 0.5 molar during incubation inhibited photosynthesis in the controls by 14% or less. This same protocol resulted in an inhibition of photosynthesis by as much as 41% in K(+)-deficient tissue. In contrast to the data obtained with leaf slices, dehydration inhibition of isolated chloroplast photosynthesis was not affected by K(+) status of parent plant material. These data are consistent with the hypothesis that one effect of leaf K(+) deficiencies on photosynthetic response to dehydration may be mediated by extra-choloroplastic factors.Ammonium ions, which facilitate stromal alkalinization, reversed the increased sensitivity of K(+)-deficient leaf slice photosynthesis to cell dehydration. However, NH(4) (+) had no effect on photosynthesis of K(+)-deficient leaf slices under nonhypertonic conditions. These data suggest that endogenous extra-chloroplastic K(+) may modulate dehydration inhibition of photosynthesis, possibly by facilitating stromal alkalinization. | description |
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| Berkowitz, G A, Whalen, C | authors |
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| Temperature Dependence of Carbon Isotope Fractionation in CAM Plants. | name |
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| The carbon isotope fractionation associated with nocturnal malic acid synthesis in Kalanchoë daigremontiana and Bryophyllum tubiflorum was calculated from the isotopic composition of carbon-4 of malic acid, after appropriate corrections. In the lowest temperature treatment (17 degrees C nights, 23 degrees C days), the isotope fractionation for both plants is -4 per thousand (that is, malate is enriched in (13)C relative to the atmosphere). For K. daigremontiana, the isotope fractionation decreases with increasing temperature, becoming approximately 0 per thousand at 27 degrees C/33 degrees C. Detailed analysis of temperature effects on the isotope fractionation indicates that stomatal aperture decreases with increasing temperature and carboxylation capacity increases. For B. tubiflorum, the temperature dependence of the isotope fractionation is smaller and is principally attributed to the normal temperature dependences of the rates of diffusion and carboxylation steps. The small change in the isotopic composition of remaining malic acid in both species which is observed during deacidification indicates that malate release, rather than decarboxylation, is rate limiting in the deacidification process. | description |
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| Deleens, E, Treichel, I, O'leary, M H | authors |
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| Localization, purification, and characterization of shikimate oxidoreductase-dehydroquinate hydrolyase from stroma of spinach chloroplasts. | name |
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| The stroma of chloroplasts is probably the sole site of the shikimate pathway enzymes shikimate oxidoreductase/dehydroquinate hydrolyase (SORase/DHQase) in spinach leaves. (a) The chromatographic behavior of the bifunctional protein SORase/DHQase on several separation materials with extracts from stroma compared with leaf extracts showed only one peak of enzymic activity originating from the stroma. (b) Polyacrylamide gel electrophoresis (PAGE) of these extracts followed by specific staining resulted in the same pattern without a band of extraplastidic enzyme. (c) In protoplast fractionation experiments it was shown that SORase/DHQase was present only in the soluble chloroplast protein fraction.An improved purification procedure for SORase/DHQase from stroma of chloroplasts, yield 40%, 1600 times as pure, gave essentially one protein band on sodium dodecyl sulfate-PAGE. Our results demonstrate that both enzyme functions are carried out by a single polypeptide. Nondenaturing PAGE exhibited a pattern of four bands with SORase/DHQase showing that they differ in charge but not in their molecular weight. Molecular weight was determined to be 67 kilodaltons (gel filtration) and 59 kilodaltons (PAGE) for all four forms. It was proven they were not due to artifacts. The four forms show similar kinetic properties, their K(m) and pH optima differing only very slightly. Response to some metabolites is reported. | description |
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| Fiedler, E, Schultz, G | authors |
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| Phosphatidylglycerol synthesis in pea chloroplasts: pathway and localization. | name |
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| Isolated intact pea chloroplasts synthesized phosphatidylglycerol from either [(14)C]acetate or [(14)C]glycerol 3-phosphate. Both time-course and pulse-chase labeling studies demonstrated a precursor-product relationship between newly synthesized phosphatidic acid and newly synthesized phosphatidylglycerol.The synthesis both of CDP-diacylglycerol from exogenous phosphatidic acid and CTP, and of phosphatidylglycerol from exogenous CDP-diacylglycerol and glycerol 3-phosphate, could be assayed in fractions obtained from disrupted chloroplasts. Moreover, the enzymes catalyzing these reactions were localized in the inner envelope membrane. Exogenous phosphatidic acid was incorporated into phosphatidylglycerol, but only following its incorporation into CDP-diacylglycerol. Finally, radio-active phosphatidic acid synthesized in the envelope membranes from [(14)C]palmitoyl-ACP and 1-oleoyl-glycerol 3-phosphate was sequentially incorporated into labeled CDP-diacylglycerol and phosphatidylglycerol upon the addition of appropriate substrates and cofactors. Thus, we have demonstrated that (a) the synthesis of phosphatidylglycerol in chloroplasts occurs by the pathway: phosphatidic acid --> CDP-diacylglycerol -->--> phosphatidylglycerol, and (b) phosphatidylglycerol synthesis is located in the inner envelope membrane. | description |
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| Andrews, J, Mudd, J B | authors |
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| Proteolytic activity in anther extracts of fertile and cytoplasmic male sterile petunia. | name |
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| Proteolytic activity is compared in anther extracts from Petunia parodii fertile and cytoplasmic male sterile lines. It is characterized relative to developmental stage of the anthers, effect of variable incubation times, pH of isolation buffers, and degradation of marker proteins. In fertile anthers, proteolytic activity increases at the end of microsporogenesis and peaks early in microgametogenesis. Degradation is most severe in extracts of fertile anthers and in high molecular weight proteins and reaches its maximum within 20 minutes. Degradation of marker proteins is greatest at pH 5.6 to 8.0 in fertile anther extracts and is eliminated under strong acid conditions (pH 2.8 to 4.0) in both fertile and cytoplasmic male sterile anther extracts. Marker proteins degrade more severely in extracts of fertile anthers; however, the order of substrate sensitivity-myosin > phosphorylase b > bovine serum albumin and ovalbumin > beta-galactosidase-is the same in extracts from fertile and cytoplasmic male sterile anthers. | description |
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| Wu, F S, Murry, L E | authors |
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| Reduced phytic Acid content does not have an adverse effect on germination of soybean seeds. | name |
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| Altering the level of phytic acid phosphorus by nutritional means had no effect on the ability of soybean (Glycine max L. [Merr.], cv ;Williams 79') seeds to germinate under laboratory or greenhouse conditions. Dry matter moved out of the cotyledons at similar rates whether the germinating seeds initially contained low (0.19), medium (0.59), or high (1.00 milligram per seed) phytic acid phosphorus. Growth of roots and shoots from 3 to 9 days after planting was similar for seeds containing low and medium levels of phytic acid phosphorus. The medium level of phytic acid resembles that found in field-grown seed, so it is clear that soybean seeds normally contain a phosphorus reserve far above that needed for germination and early seedling growth. | description |
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| Raboy, V, Hudson, S J, Dickson, D B | authors |
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| Effect of Triacontanol on Chlamydomonas: I. Stimulation of Growth and Photosynthetic CO(2) Assimilation. | name |
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| Treatment of Chlamydomonas reinhardtii cells, cultured at 5% CO(2), with 1 to 1000 micrograms triacontanol (TRIA) per liter resulted in 21 to 35% increases in cell density, 7 to 31% increases in total chlorophyll, and 20 to 100% increases in photosynthetic CO(2) assimilation. The increase in CO(2) fixation with TRIA treatment occurred before, and was independent of, increases in total chlorophyll or cell number. Chlamydomonas cells responded to a broad range of TRIA concentrations that were at least one order of magnitude above the optimum concentration established for higher plants. The necessity for larger concentrations of TRIA may be due to destabilizing effects of Ca(2+) and K(+) present in the Chlamydomonas growth medium. These ions caused flocculation of the colloidally dispersed TRIA in apparent competition with binding of [(14)C]TRIA to Chlamydomonas cells. Octacosanol inhibited the effect of TRIA on photosynthetic CO(2) assimilation. TRIA treatment did not alter the distribution of (14)C-label among photosynthetic products. The effect of TRIA on photosynthetic CO(2) assimilation increased with time after treatment up to 3 days. Chlamydomonas cells that had been grown at low-CO(2) (air) did not respond to TRIA, and transfer of high-CO(2) (5%) grown cells that had responded to TRIA to a low-CO(2) atmosphere resulted in a loss of the effect of TRIA. The effect of pH on photosynthetic CO(2) assimilation indicated that CO(2) is probably the species of inorganic carbon utilized by control and TRIA-treated Chlamydomonas cells. | description |
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| Houtz, R L, Ries, S K, Tolbert, N E | authors |
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| Effect of Temperature, Oxygen, and Gibberellic Acid on the Development of Photosensitivity in Oldenlandia corymbosa L. Seeds during Their Incubation in Darkness. | name |
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| Two successive phases can be distinguished in the development of the responsiveness to light in Oldenlandia corymbosa L. seeds during their incubation in darkness. During phase I, the responsiveness to light increases with time if there is sufficient O(2), and the higher the temperature, the faster the increase. This phase is stimulated by gibberellic acid. During the following phase (II), seeds remain responsive to light at 10 or 20 degrees C, but lose their responsiveness at higher temperature (>/=30 degrees C). This second phase depends on O(2): loss of responsiveness is accelerated at lower O(2) concentration. Phase II is only slightly affected by gibberellic acid. The results are discussed in terms of variation of phytochrome and of a reaction along the transduction chain initiated by phototransformation of this pigment, which is finally expressed in germination. | description |
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| Corbineau, F, Côme, D | authors |
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| The Effect of Sugars on the Binding of [Hg]-p-Chloromercuribenzenesulfonic Acid to Leaf Tissues. | name |
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| Replacement of mannitol with sucrose decreases the binding of [(203)Hg]-p-chloromercuribenzenesulphonic acid (PCMBS) to Vicia faba leaf discs without epidermis. This decrease is optimal for 20 minutes on incubation, is concentration-dependent, and is also found with maltose and raffinose. In parallel experiments, the addition of sucrose, maltose, and raffinose during PCMBS pretreatment was shown to increase subsequent uptake of [U-(14)C]sucrose. In contrast, d- or l-glucose, 3-O-methylglucose, galactose, fructose, palatinose, turanose, or melibiose had no effect either on PCMBS binding or on [(14)C]sucrose uptake. The sucrose-induced decrease of PCMBS binding is retained after a cold and ionic shock. Measurements of specific activities of membrane fractions prepared from tissues incubated in labeled PCMBS show that the decrease concerns the 120,000 gravity pellet, but that very mild procedures must be chosen to prevent redistribution of label in the supernatant. Altogether, the data provide new support to the hypothesis that the active site of the sucrose carrier contains a group sensitive to PCMBS. | description |
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| M'batchi, B, Pichelin, D, Delrot, S | authors |
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| A simple procedure to overcome polyethelene glycol toxicity on whole plants. | name |
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| A procedure is described that can be used to minimize toxic effects of polyethylene glycol (PEG) to plants. The procedure is based on recycling nutrient solutions containing PEG-6000 through two plant cultures. Tomato plants grown in -0.3 megapascals PEG solutions used after two growth cycles exhibited minimal toxic effects. Long-term responses like dry matter production and chlorophyll content as well as short-term responses like CO(2) fixation rates and leaf conductance were severely inhibited by fresh PEG-6000 and only slightly reduced by recycled PEG-6000. Complete osmotic adjustment was obtained with tomatoes grown in recycled but not in fresh PEG solutions. | description |
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| Plaut, Z, Federman, E | authors |
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| High efficiency transformation of cultured tobacco cells. | name |
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| Tobacco calli were transformed at levels up to 50% by cocultivation of tobacco cultured cells with Agrobacterium tumefaciens harboring the binary transfer-DNA vector, pGA472, containing a kanamycin resistance marker. Transformation frequency was dependent on the physiological state of the tobacco cells, the nature of Agrobacterium strain and, less so, on the expression of the vir genes of the tumor-inducing plasmid. Maximum transformation frequency was obtained with exponentially growing plant cells, suggesting that rapid growth of plant cells is an essental factor for efficient transformation of higher plants. | description |
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| An, G | authors |
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| Effects of Nitrate Application on Amaranthus powellii Wats. : I. Changes in Photosynthesis, Growth Rates, and Leaf Area. | name |
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| Physiological effects of different nitrate applications were studied using the C(4) plant, Amaranthus powellii Wats. Plants were grown in a controlled environment chamber and watered daily with nutrient solutions containing 45, 10, 5, or 1 millimolar nitrate. Chloride and sulfate were used to keep the cation and phosphate concentrations constant. Total leaf nitrogen concentration, chlorophyll concentration, specific leaf mass, leaf area, relative growth rate, relative leaf growth rate, unit leaf rate (increase of dry mass per unit leaf area per day), net photosynthetic rate, and incident quantum yield decreased with decreasing nitrate concentration. The per cent decrease of unit leaf rate was similar to the decrease of light-saturated net photosynthetic rate; however, the decrease in relative growth rate was less than that of unit leaf rate because leaf area ratio (leaf area per unit dry mass) increased with decreasing nitrate concentration. Essential mineral concentrations per unit leaf area were about equal among all treatments. Leaf expansion, determined by stomatal density, decreased except for the 1 millimolar treatment which showed relatively more cell expansion but less cell division. Decreased nitrate application was correlated with higher osmotic potentials and lower pressure potentials (determined by pressure-volume curves), whereas leaf water potentials were equal among treatments. Even though total leaf area and shoot mass decreased with decreasing applied nitrate, the increase of the leaf area ratio may be related to selection for the highest possible growth rate. | description |
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| Hunt, E R, Weber, J A, Gates, D M | authors |
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| Role of ethylene in the senescence of isolated hibiscus petals. | name |
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| Senescence of petals isolated from flowers of Hibiscus rosa-sinensis L. (cv Pink Versicolor) was associated with increased ethylene production. Exposure to ethylene (10 microliters per liter) accelerated the onset of senescence, as indicated by petal in-rolling, and stimulated ethylene production. Senescence was also hastened by basal application of 1-aminocyclopropane-1-carboxylic acid (ACC). Aminooxyacetic acid, an inhibitor of ethylene biosynthesis, effectively inhibited ethylene production by petals and delayed petal in-rolling. In marked contrast to these results with mature petals, immature petals isolated from flowers the day before flower opening did not respond to ethylene in terms of an increase in ethylene production or petal in-rolling. Furthermore, treatment with silver thiosulfate the day before flower opening effectively prevented petal senescence, while silver thiosulfate treatment on the morning of flower opening was ineffective. Application of ACC to both immature and mature petals greatly stimulated ethylene production indicating the presence of an active ethylene-forming enzyme in both tissues. Immature petals contained less free ACC than mature, presenescent petals and appeared to possess a more active system for converting ACC into its conjugated form. Thus, while the nature of the lack of responsiveness of immature petals to ethylene is unknown, ethylene production in hibiscus petals appears to be regulated by the control over ACC availability. | description |
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| Woodson, W R, Hanchey, S H, Chisholm, D N | authors |
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| Properties of the Mg-Protoporphyrin IX Monomethyl Ester (Oxidative) Cyclase System. | name |
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| Mg-protoporphyrin IX monomethyl ester (oxidative) cyclase, the enzyme system responsible for the formation of the chlorophyll isocyclic ring, exhibits requirements for both essential sulfhydryls and essential disulfides. It is inhibited by N-ethylmaleimide, dithiothreitol, and beta-mercaptoethanol, but not by sodium arsenite. This enzyme system shows some substrate specificity: (a) the 6-side-chain of the macrocycle can either be a methyl propionate ester, or its beta-hydroxy or beta-keto derivatives; (b) the 7-side-chain can either be a propionic acid or a methyl propionate ester; (c) both the 4-vinyl and the 4-ethyl series can serve as substrates, at least at the beta-keto ester level; (d) the activity appears to be lost if the side-chain in the 2-position is reduced from a vinyl to an ethyl. | description |
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| Wong, Y S, Castelfranco, P A | authors |
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| 3-Deoxy-d-arabino-Heptulosonate 7-Phosphate Synthase from Carrot Root (Daucus carota) Is a Hysteretic Enzyme. | name |
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| Roots of carrots (Daucus carota) contain three activities of 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthase, the enzyme that catalyzes the first step of the shikimate pathway. The three activities, enzymes I, II, and III, are separated by chromatography on phosphocellulose. Enzyme III, purified to electrophoretic homogeneity, has a native molecular weight of 103,000 and consists of two identical subunits of 53,000 daltons each. Double reciprocal plots of reaction velocity versus substrate concentration yield K(m) values of 0.03 and 0.07 millimolar for P-enolpyruvate and erythrose-4-P, respectively. Both products, DAHP and orthophosphate, inhibit the enzyme. Enzyme III is a hysteretic enzyme that is activated by physiological concentrations of l-tryptophan and Mn(2+), both of which also partially eliminate the hysteretic lag. Feedback activation of carrot DAHP synthase by tryptophan is interpreted to be an early regulatory signal for polyphenol biosynthesis. The three carrot DAHP synthase isoenzymes share antigenic determinants. | description |
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| Suzich, J A, Dean, J F, Herrmann, K M | authors |
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| Studies on the photoactivation of the water-oxidizing enzyme : I. Processes limiting photoactivation in hydroxylamine-extracted leaf segments. | name |
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| In weak yet optimal light intensity, complete photoactivation of the water-oxidizing enzyme in NH(2)OH-extracted wheat (Triticum aestivum, var Oasis) leaf segments could be obtained only after long dark preincubation. Photoactivation was not affected by ethylenediaminetetraacetate or inhibitors of photophosphorylation and protein synthesis, but was partially inhibited by a divalent cation ionophore. Complete photoactivation required ligation of approximately 4 Mn by the water oxidizing enzyme.WITHOUT DARK PREINCUBATION, PHOTOSYSTEM II (PSII) WAS SUSCEPTIBLE TO WEAK LIGHT PHOTOINHIBITION RESULTING IN: (a) 50% maximum decrease in photooxidation of artificial electron donors by PSII: (b) increased times for the variable fluorescence rise (with 3-(3,4-dichlorophenyl)-1,1-dimethyl urea): (c) abolishment of photoactivation: and (d) the imposition of sensitivity to inhibitors of photophosphorylation and 70S but not 80S protein synthesis on subsequent light-dependent recovery from photoinhibition and recovery of O(2) evolution. Decrease in susceptibility to photoinhibition and increase in rates of photoactivation resulting from dark preincubations proved closely correlated. Neither protein synthesis nor increases in abundances of thylakoid Mn(2+) and Ca(2+) were required for escape from photoinhibition. However, photoactivation of the wateroxidizing enzyme in NH(2)OH-extracted Chlamydomonas occurred in absence of dark preincubation and protein synthesis. Results are discussed in the context of disassembly/reassembly/resynthesis of specific PSII polypeptides. | description |
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| Callahan, F E, Cheniae, G M | authors |
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| Enzymic degradation of allantoate in developing soybeans. | name |
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| A Mn(2+)-dependent enzymic breakdown of allantoate has been detected in crude and partially purified extracts of developing soybeans. The products detected were CO(2), NH(3), glyoxylate, labile glyoxylate derivatives, and low levels of urea. Urea is initially produced at less than 10% the rate of urease-independent CO(2) release indicating that the activity is not allantoate amidinohydrolase (i.e. urea is not directly cleaved off allantoate). The urease-independent CO(2) releasing activity has an apparent K(m) of 1.0 millimolar for allantoate. Ethylenediaminetetraacetate, borate, and acetohydroxamate (all at 10 millimolar) inhibit the enzymic production of NH(3), CO(2), and labile glyoxylate derivatives from allantoate. However, the potent urease inhibitor, phenyl phosphordiamidate does not inhibit CO(2) and NH(3) release indicating that the action of acetohydroxamate is not due to its inhibition of urease. That the allantoatedegrading activity was more than 5-fold greater in seed coats than in embryos is consistent with the data of Rainbird et al. (Plant Physiol 1984 74: 329-334) which indicate that available ureides are metabolized before reaching the embryo. 2-Ethanolthio, 2'ureido, acetic acid (NH(2)COHNCHCO(2)HSCH(2)CH(2)OH), the first allantoate-derived product detected by HPLC analysis, is an addition produced of mercaptoethanol with an unidentified enzymically produced ureido intermediate that is not derived from ureidoglycolate or oxalurate. | description |
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| Winkler, R G, Polacco, J C, Blevins, D G, Randall, D D | authors |
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| Peroxidases and glycosidases in intercellular fluids from noninoculated and rust-affected wheat leaves : isozyme assay on nitrocellulose blots from two-dimensional gels. | name |
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| Proteins in intercellular washing fluid (IWF) from noninoculated and stem rust-affected wheat leaves were separated by isoelectric focusing and polyacrylamide gel electrophoresis under nondenaturing conditions, transferred to nitrocellulose membranes, and assayed in situ for peroxidase and glycosidase activity.Two infection-related peroxidase isozymes were detected in addition to more than ten that were present only in noninoculated plants. One other peroxidase isozyme was present in much higher concentration in IWF from infected leaves than in IWF from noninoculated leaves.IWF contained many polymorphic glycosidases. A new method is described to localize the glycosidase isozymes accurately on nitrocellulose blots for evaluation of their substrate specificities: each blot was developed with the appropriate p-nitrophenyl glycoside to reveal glycosidase activity, then reprobed for concanavalin A-binding glycoproteins to serve as an internal reference frame for blot-to-blot comparisons. This procedure also provided information on glycosylation of the isozymes.The locations of at least 15 (out of 37) isozymes were coincident with concanavalin A binding, including those of all 10 alpha-d-mannosidases, and of 6 beta-d-xylosidases. On five areas of the blots there was coincidence of beta-d-xylosidase and alpha-l-arabinosidase activity. Three of these areas corresponded to three of the most prominent Coomassie brilliant blue-stainable IWF proteins in gels. Isozymes that could convert p-nitrophenyl-beta-d-glucoside, -beta-d-galactoside, and/or -beta-d-fucoside revealed a complex pattern of partially overlapping substrate specificities: two isozymes utilized both glucose and fucose derivatives, one utilized all three derivatives, and several others converted only one of the three substrates. No enzymes were detected with activity on p-nitrophenyl-beta-d-galactosaminide, -beta-l-fucoside, or -alpha-d-galactoside. No additional glycosidases were detected in IWF from stem rust-affected leaves. | description |
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| Holden, D W, Rohringer, R | authors |
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| Light Activation of NADP-Malate Dehydrogenase in Guard Cell Protoplasts from Vicia faba L. | name |
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| Light-induced swelling of guard cell protoplasts (GCP) from Vicia faba was accompanied by increases in content of K(+) and malate. DCMU inhibited the increase of K(+) and malate, and consequently swelling.Effect of light on the activity of selected enzymes that take part in malate formation was studied. When isolated GCP were illuminated, NADP-malate dehydrogenase (NADP-MDH) was activated, and the activity reached a maximum within 5 minutes. The enzyme activity underwent 5- to 6-fold increase in the light. Upon turning off the light, the enzyme was inactivated in 5 minutes NAD-MDH and phosphoenolpyruvate carboxylase (PEPC) were not influenced by light. The rapid light activation of NADP-MDH was inhibited by DCMU, suggesting that the enzyme was activated by reductants from the linear electron transport in chloroplasts. An enzyme localization study by differential centrifugation indicates that NADP-MDH is located in the chloroplasts, NAD-MDH in the cytosol and mitochondria, and PEPC in the cytosol. After light activation, the activity of NADP-MDH in guard cells was 10 times that in mesophyll cells on a chlorophyll basis. The physiological significance of light-dependent activation of NADP-MDH in guard cells is discussed in relation to stomatal movement. | description |
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| Gotow, K, Tanaka, K, Kondo, N, Kobayashi, K, Syōno, K | authors |
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| Photosynthetic Responses to Dynamic Light Environments by Hawaiian Trees : Time Course of CO(2) Uptake and Carbon Gain during Sunflecks. | name |
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| Gas exchange responses to rapid changes in light were studied in a C(3) tree, Claoxylon sandwicense Muell-Arg and a C(4) tree, Euphorbia forbesii Sherff that are native to the understory of a mesic Hawaiian forest. When light was increased to 500 micromoles per meter per second following a 2 hour preexposure at 22 micromoles per meter per second, net CO(2) uptake rates and stomatal conductance gradually increased for over 1 hour in C. sandwicense but reached maximum values within 30 minutes in E. forbesii. Calculation of the intercellular CO(2) pressures indicated that the primary limitation to CO(2) uptake during this induction was nonstomatal in both species. The photosynthetic response to simulated sunflecks (lightflecks) was strongly dependent on the induction state of the leaf. Total CO(2) uptake during a lightfleck was greater and the response was faster after exposure of the leaf to high light than when the leaf had been exposed only to low light for the previous 2 hours. During a series of lightflecks, induction resulted in increased CO(2) uptake in successive lightflecks. Significant postillumination CO(2) fixation was evident and contributed substantially to the total carbon gain, especially for lightflecks of 5 to 20 seconds' duration. | description |
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| Pearcy, R W, Osteryoung, K, Calkin, H W | authors |
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| An investigation into the role of photosynthesis in regulating ATP levels and rates of h efflux in isolated meosphyll cells. | name |
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| Aerated and stirred 10-ml suspensions of mechanically isolated Asparagus sprengeri Regel mesophyll cells were used for simultaneous measurements of net H(+) efflux and steady-state ATP levels.Initial rates of medium acidification indicated values for H(+) efflux in the light and dark of 0.66 and 0.77 nanomoles H(+)/10(6) cells per minute, respectively. When the medium pH was maintained at 6.5, with a pH-stat apparatus, rates of H(+) efflux remained constant. Darkness or DCMU, however, stimulated H(+) efflux by 100% or more. Darkness increased ATP levels by 33% and a switch from dark to light reduced ATP levels by 31%. In the absence of aeration, illumination prevented the accumulation of respiratory CO(2) and the buffering capacity of the medium was about 50% less than that found in the nonilluminated nonaerated medium. As a result, rates of pH decline were similar even though the dark rate of H(+) efflux was approximately 50% greater.Proposals that photosynthesis stimulates H(+) efflux are based on changes in the rate of pH decline. The present data indicate that photosynthesis inhibits H(+) efflux and that changes in rates of pH decline should not be equated with changes in the rate of H(+) efflux. | description |
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| Bown, A W, Nicholls, F | authors |
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| Biosynthesis of alpha-Amylase in Vigna mungo Cotyledon. | name |
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| In vitro translation of RNA extracted from Vigna mungo cotyledons showed that alpha-amylase is synthesized as a polypeptide with a molecular mass of 45,000, while cotyledons contain a form of alpha-amylase with a molecular mass of 43,000. To find out whether the 45,000 molecular mass polypeptide is a precursor to the 43,000 found in vivo, the cell free translation systems were supplemented with canine microsomal membrane; when mRNA was translated in the wheat germ system supplemented with canine microsomes, the 45,000 molecular mass form was not processed to a smaller form but the precursor form was partly processed in the membrane-supplemented reticulocyte lysate system. When V. mungo RNA was translated in Xenopus oocyte system, only the smaller form (molecular mass 43,000) was detected. Involvement of contranslational glycosylation in the maturating process of the alpha-amylase was ruled out because there was no effect of tunicamycin, and the polypeptide was resistant to endo-beta-H or endo-beta-D digestion. We interpret these results to mean that the 45,000 molecular mass form is a precursor with a signal peptide or transit sequence, and that the 43,000 molecular mass is the mature form of the protein. | description |
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| Tomura, H, Koshiba, T | authors |
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| Interspecific Variation in SO(2) Flux : Leaf Surface versus Internal Flux, and Components of Leaf Conductance. | name |
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| The objective of this study was to clarify the relationships among stomatal, residual, and epidermal conductances in determining the flux of SO(2) air pollution to leaves. Variations in leaf SO(2) and H(2)O vapor fluxes were determined using four plant species: Pisum sativum L. (garden pea), Lycopersicon esculentum Mill. flacca (mutant of tomato), Geranium carolinianum L. (wild geranium), and Diplacus aurantiacus (Curtis) Jeps. (a native California shrub). Fluxes were measured using the mass-balance approach during exposure to 4.56 micromoles per cubic meter (0.11 microliters per liter) SO(2) for 2 hours in a controlled environmental chamber. Flux through adaxial and abaxial leaf surfaces with closed stomata ranged from 1.9 to 9.4 nanomoles per square meter per second for SO(2), and 0.3 to 1.3 millimoles per square meter per second for H(2)O vapor. Flux of SO(2) into leaves through stomata ranged from approximately 0 to 8.5 (dark) and 3.8 to 16.0 (light) millimoles per square meter per second. Flux of H(2)O vapor from leaves through stomata ranged from approximately 0 to 0.6 (dark) to 0.4 to 0.9 (light) millimole per square meter per second. Lycopersicon had internal flux rates for both SO(2) and H(2)O vapor over twice as high as for the other species. Stomatal conductance based on H(2)O vapor flux averaged from 0.07 to 0.13 mole per square meter per second among the four species. Internal conductance of SO(2) as calculated from SO(2) flux was from 0.04 mole per square meter per second lower to 0.06 mole per square meter per second higher than stomatal conductance. For Pisum, Geranium, and Diplacus stomatal conductance was the same or slightly higher than internal conductance, indicating that, in general, SO(2) flux could be predicted from stomatal conductance for H(2)O vapor. However, for the Lycopersicon mutant, internal leaf conductance was much higher than stomatal conductance, indicating that factors inside leaves can play a significant role in determining SO(2) flux. | description |
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| Olszyk, D M, Tingey, D T | authors |
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| Interaction of wheat germ ca-dependent protein kinases with calmodulin antagonists and polyamines. | name |
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| The two soluble Ca(2+)-dependent protein kinases resolved from wheat (Triticum aestivum) embryo (protein kinases I and II) are inhibited by the phenothiazine-derived calmodulin antagonists trifluoperazine fluphenazine, and chlorpromazine. Protein kinases I and II are also inhibited by a variety of other calmodulin antagonists (including calmidazolium, amitriptyline, and iprindole), phosphodiesterase inhibitors (including flufenamic acid and papavarine) and by lanthanides. A number of compounds that inhibit mammalian Ca(2+) - and phospholipid-activated protein kinase (protein kinase C) including quercetin, polymixin B sulfate, and polyamines (as well as phenothiazine derivatives) also inhibit protein kinases I and II. Poly-l-lysine and poly-l-ornithine activate both plant Ca(2+)-dependent protein kinases. | description |
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| Polya, G M, Micucci, V | authors |
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| A role for fructose 2,6-bisphosphate in regulating carbohydrate metabolism in guard cells. | name |
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| Fructose 2,6-bisphosphate (Fru2,6P(2)) appears to function as a regulator metabolite in glycolysis and gluconeogenesis in animal tissues, yeast, and the photosynthetic cells of leaves. We have investigated the role of Fru2,6P(2) in guard-cell protoplasts from Vicia faba L. and Pisum sativum L. (Argenteum mutant), and in epidermal strips purified by sonication from all cells except for the guard cells. Guard-cell protoplasts were separated into fractions enriched in cytosol and in chloroplasts by passing them through a nylon net, followed by silicone oil centrifugation. The cytosol contained a pyrophosphate: fructose 6-phosphate phosphotransferase (involved in glycolysis) which was strongly stimulated by Fru2,6P(2). A cytosolic fructose 1,6-bisphosphatase (a catalyst of gluconeogenesis) was inhibited by Fru2,6P(2). There was virtually no fructose 1,6-bisphosphatase activity in guard-cell chloroplasts of V. faba. It is therefore unlikely that the starch formed in these chloroplasts originates from imported triose phosphates or phosphoglycerate.The level of Fru2,6P(2) in guard-cell protoplasts and epidermal strips was about 0.1 to 1 attomole per guard cell in the dark (corresponding to 0.05 to 0.5 nanomole per milligram chlorophyll) and increased three- to tenfold within 15 minutes in the light. Within the same time span, hexose phosphate levels in guard-cell protoplasts declined to approximately one-half, indicating that acceleration of glycolysis involved stimulation of reactions using hexose phosphates. The level of Fru2,6P(2) in guard cells appears to determine the direction in which carbohydrate metabolism proceeds. | description |
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| Hedrich, R, Raschke, K, Stitt, M | authors |
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| Identification and Isolation of Single Cells that Produce Somatic Embryos at a High Frequency in a Carrot Suspension Culture. | name |
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| A system was established in which single cells differentiated to embryos at a high frequency. Small spherical single cells from a carrot (Daucus carota L. cv "Kurodagosun") cell suspension culture were obtained by fractionation through sieving, using nylon screens and then density gradient centrifugation in Percoll solutions. Eighty-five to 90% of these small single cells differentiated to embryos when they were cultured in a medium containing 2,4-dichlorophenoxyacetic acid (5 x 10(-8) molar), zeatin (10(-6) molar), and mannitol (0.2 molar) for 7 days, followed by transfer to a medium containing zeatin (10(-7) molar) but no auxin. This indicates that there are at least two phases in the differentiation of embryos from single cells. The progression of the first phase required exogenous auxin, whereas that of the second phase was inhibited by the same growth regulator. The relationship between the morphology and potency for embryogenesis from single cells was discussed. The system established here is a useful one for investigation of differentiation process from a single cell to a whole plant via embryogenesis, especially in its early stage. | description |
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| Nomura, K, Komamine, A | authors |
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| Photoinhibition and Reactivation of Photosynthesis in the Cyanobacterium Anacystis nidulans. | name |
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| The susceptibility of photosynthesis to photoinhibition and its recovery were studied on cultures of the cyanobacterium Anacystis nidulans. Oxygen evolution and low temperature fluorescence kinetics were measured. Upon exposure to high light A. nidulans showed a rapid decrease in oxygen evolution followed by a quasi steady state rate of photosynthesis. This quasi steady state rate decreased with increasing photon flux density of the photoinhibitory light. Reactivation of photosynthesis in dim light after the photoinhibitory treatment was rapid: 85 to 95% recovery occurred within 2 hours. In the presence of the translation inhibitor, streptomycin (250 micrograms per milliliter), no reactivation occurred. We also found that the damage increased dramatically if the high light treatment was done with streptomycin added. A transcription inhibitor, rifampicin, did not inhibit the reactivation process. Based on these data we conclude that the photoinhibitory damage observed is the net result of a balance between the photoinhibitory process and the operation of the repairing mechanism(s). | description |
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| Samuelsson, G, Lönneborg, A, Rosenqvist, E, Gustafsson, P, Oquist, G | authors |
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| Role of Ca and EGTA on Stomatal Movements in Commelina communis L. | name |
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| Ca(2+) (0.1-1.0 millimolar) accelerated dark-induced stomatal closure and reduced stomatal apertures in the light in epidermal peels of Commelina communis L. In contrast, ethyleneglycol-bis-(beta-aminoethyl ether) N,N'tetraacetic acid (EGTA) (2 millimolar), a Ca(2+) chelator, prevented closure in the dark and accelerated opening in the light. EGTA did not promote significant opening in the dark. It is therefore concluded that EGTA does not increase ion uptake into guard cells, but rather prevents ion efflux. Addition of EGTA to incubating solutions with 10 millimolar KCl resulted in steady state apertures of 15.6 micrometers, whereas in the absence of EGTA similar apertures required 55 millimolar KCl and 150 millimolar KCl was needed in the presence of 1 millimolar CaCl(2). The results demonstrate the importance of Ca(2+) in the regulation of stomatal closure and point to a role of Ca(2+) in the regulation of K(+) efflux from stomatal guard cells. | description |
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| Schwartz, A | authors |
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| Photocontrol of Hypocotyl Elongation in Light-Grown Cucumis sativus L. : Responses to Phytochrome Photostationary State and Fluence Rate. | name |
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| The effects of the calculated photostationary state of phytochrome (phi(c)) and the photon fluence rate on the elongation growth of the hypocotyl of light-grown seedlings of Cucumis sativus L. are examined. Two threshold responses to phi(c) are found at values of 0.06 and 0.43. At phi(c) = 0.06, there is no response at any fluence rate. In the phi(c) range 0.1 to 0.43, elongation growth does not respond to changes in phi(c). Above the second threshold (phi(c) = 0.43), there is a strong response to changes in phi(c). At all values of phi(c) at and above 0.1, there is a response to fluence rate. A linear relationship can be demonstrated between a factor comprised of the logarithm of phytochrome cycling rate (a fluence-rate-dependent process) and phi(c), and the growth response. | description |
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| Gaba, V, Black, M | authors |
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| Factors affecting ice nucleation in plant tissues. | name |
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| Factors affecting the ice nucleation temperature of plants and plant tissues were examined. The mass of a sample had a marked effect on ice nucleation temperature. Small tissue samples supercooled to -10 degrees C and were not accurate predictors of the nucleation temperature of intact plants in either laboratory or field experiments. This effect was not unique to plant tissues and was observed in autoclaved and control soil samples. Ice nucleation temperatures of bean, corn, cotton, and soybean seedlings were influenced by the length of subzero exposure, presence of ice nucleation active bacteria, and leaf surface wetness. The number of factors influencing ice nucleation temperature suggested that predicting the freezing behavior of plants in the field will be complex. | description |
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| Ashworth, E N, Davis, G A, Anderson, J A | authors |
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