bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of a Light-Controlled Anion Channel in the Plasma Membrane of Mesophyll Cells of Pea
Authors: Elzenga JTM. , Van Volkenburgh E. 
Published: October 2016

Abstract
In leaf mesophyll cells of pea (Pisum sativum) light induces a transient depolarization that is at least partly due to an increased plasma membrane conductance for anions. Several channel types were identified in the plasma membrane of protoplasts from mesophyll cells using the patch-clamp technique. One of these was an anion channel with a single-channel conductance of 32 picasiemens in symmetrical 100/100 KCl solutions. In asymmetrical solutions the reversal potential indicates a high selectivity for Cl- over K+ at high cytoplasmic Cl-. At negative membrane voltages the channel openings were interrupted by very short closures. In the open channel conductance several substrates were identified. At a cytoplasmic negative logarithm of Ca concentration higher than 6.3, no channel openings were observed. When the protoplast was illuminated in the cell-attached configuration, at least one channel type had a higher opening probability. This channel can tentatively be identified as the above-described anion channel based on conductance and the characteristic short closures at negative membrane potentials. This light activation of the 32-picasiemen anion channel is a strong indication that this channel conducts the light-induced depolarizing current. Because channel activity is strongly Ca2+-dependent, a role of cytoplasmic Ca2+ concentration changes in the light activation of the conductance is discussed.
Glutathione-Mediated Regulation of ATP Sulfurylase Activity, SO42- Uptake, and Oxidative Stress Response in Intact Canola Roots
Authors: Lappartient A. G. , Touraine B. 
Published: October 2016

Abstract
The dual role of glutathione as a transducer of S status (A.G. Lappartient and B. Touraine [1996] Plant Physiol 111: 147–157) and as an antioxidant was examined by comparing the effects of S deprivation, glutathione feeding, and H2O2 (oxidative stress) on SO42- uptake and ATP sulfurylase activity in roots of intact canola (Brassica napus L.). ATP sulfurylase activity increased and SO42- uptake rate severely decreased in roots exposed to 10 mM H2O2, whereas both increased in S-starved plants. In split-root experiments, an oxidative stress response was induced in roots remote from H2O2 exposure, as revealed by changes in the reduced glutathione (GSH) level and the GSH/oxidized glutathione (GSSG) ratio, but there was only a small decrease in SO42- uptake rate and no effect on ATP sulfurylase activity. Feeding plants with GSH increased GSH, but did not affect the GSH/GSSG ratio, and both ATP sulfurylase activity and SO42- uptake were inhibited. The responses of the H2O2-scavenging enzymes ascorbate peroxidase and glutathione reductase to S starvation, GSH treatment, and H2O2 treatment were not to glutathione-mediated S demand regulatory process. We conclude that the regulation of ATP sulfurylase activity and SO42- uptake by S demand is related to GSH rather than to the GSH/GSSG ratio, and is distinct from the oxidative stress response.
Asymmetric Distribution of Acetylcholinesterase in Gravistimulated Maize Seedlings
Authors: Momonoki Y. S. 
Published: October 2016

Abstract
Acetylcholinesterase (AChE) activity has previously been studied by this laboratory and shown to occur at the interface between the stele and cortex of the mesocotyl of maize (Zea mays L.) seedlings. In this work we studied the distribution of AChE activity in 5-d-old maize seedlings following a gravity stimulus. After the stimulus, we found an asymmetric distribution of the enzyme in the coleoptile, the coleoptile node, and the mesocotyl of the stimulated seedlings using both histochemical and colorimetric methods for measuring the hydrolysis of acetylthiocholine. The hydrolytic capability of the esterase was greater on the lower side of the horizontally placed seedlings. Using the histochemical method, we localized the hydrolytic capability in the cortical cells around the vascular stele of the tissues. The hydrolytic activity was inhibited 80 to 90% by neostigmine, an inhibitor of AChE. When neostigmine was applied to the corn kernel, the gravity response of the seedling was inhibited and no enzyme-positive spots appeared in the gravity-stimulated seedlings. We believe these results indicate a role for AChE in the gravity response of maize seedlings.
Influence of Environmental Stress on Biomass Partitioning in Transgenic Tobacco Plants Expressing the Movement Protein of Tobacco Mosaic Virus
Authors: Balachandran S. , Hull R. J. , Martins R. A. , Vaadia Y. , Lucas W. J. 
Published: October 2016

Abstract
The influence of various environmental factors on biomass partitioning between shoots and roots in transgenic tobacco (Nicotiana tabacum) plants expressing the movement protein (MP) of tobacco mosaic virus (TMV) was investigated. TMV-MP-expressing transgenic plants exhibited a root-to-shoot ratio that was approximately 40% below that of transgenic vector control plants. When transgenic plants expressing the TMV-MP were subjected to water-stress conditions, the root-to-shoot ratio was increased to a value comparable to that of control plants subjected to the same water-stress treatment. Although the root-to-shoot ratio was increased by N or P deficiencies, the TMV-MP-induced alteration in biomass partitioning was not overcome. Surprisingly, under K+-deficient growth conditions, both TMV-MP-expressing and control plants exhibited reduced root-to-shoot ratios when compared with plants grown in the presence of sufficient K+. Furthermore, plant growth under K+-deficient conditions did not alleviate the influence of the TMV-MP over resource allocation to the roots. These results are discussed in terms of possible mechanisms by which stress signals could cause an alteration in biomass partitioning between shoots and roots in control and transgenic tobacco plants expressing the TMV-MP.
Metabolic Control of Avocado Fruit Growth (Isoprenoid Growth Regulators and the Reaction Catalyzed by 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase)
Authors: Cowan A. K. , Moore-Gordon C. S. , Bertling I. , Wolstenholme B. N. 
Published: October 2016

Abstract
The effect of isoprenoid growth regulators on avocado (Persea americana Mill. cv Hass) fruit growth and mesocarp 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) activity was investigated during the course of fruit ontogeny. Both normal and small-fruit phenotypes were used to probe the interaction between the end products of isoprenoid biosynthesis and the activity of HMGR in the metabolic control of avocado fruit growth. Kinetic analysis of the changes in both cell number and size revealed that growth was limited by cell number in phenotypically small fruit. In small fruit a 70% reduction in microsomal HMGR activity was associated with an increased mesocarp abscisic acid (ABA) concentration. Application of mevastatin, a competitive inhibitor of HMGR, reduced the growth of normal fruit and increased mesocarp ABA concentration. These effects were reversed by co-treatment of fruit with mevalonic acid lactone, isopentenyladenine, or N-(2-chloro-4-pyridyl)-N-phenylurea, but were not significantly affected by either gibberellic acid or stigmasterol. However, stigmasterol appeared to partially restore fruit growth when co-injected with mevastatin in either phase II or III of fruit growth. In vivo application of ABA reduced fruit growth and mesocarp HMGR activity and accelerated fruit abscission, effects that were reversed by co-treatment with isopentenyladenine. Together, these observations indicate that ABA accumulation down-regulates mesocarp HMGR activity and fruit growth, and that in situ cytokinin biosynthesis modulates these effects during phase I of fruit ontogeny, whereas both cytokinins and sterols seem to perform this function during the later phases.
Characterization of the Cell Wall Microdomain Surrounding Plasmodesmata in Apple Fruit
Authors: Roy S. , Watada A. E. , Wergin W. P. 
Published: October 2016

Abstract
In fleshy fruits ripening is generally associated with a loss in tissue firmness resulting from depolymerization of wall components and separation of adjacent cells. In the regions of the wall that contain plasmodesmata, the usual sequences of ripening events, i.e. depolymerization of the middle lamellae and splitting of the walls, are not observed. In the present study we attempted to characterize in apple (Malus domestica Borkh.) fruit the structural microdomain of the cell wall that surrounds the plasmodesmata by in muro visualization of the cell wall components. Anionic sites of galacturonic acids were labeled with cationic gold. Low-esterified homogalacturonans were labeled with the monoclonal antibody JIM 5. In addition, a polyclonal antibody directed toward [beta](1-&gt;3)-glucopyranose was used to target callose in situ. The results indicated that the plasmodesmata-wall complexes were surrounded by a pectic microdomain. This domain was composed of low-esterified homogalacturonans that were not involved in calcium cross-bridging but were probably surrounded by a cationic environment. These structural features may result in the prevention of normal cell wall separation in regions containing plasmodesmata. However, observations by low-temperature scanning electron microscopy suggested that splitting of these walls ruptured the plasmodesmata and ultimately resulted in the spatial separation of adjacent cells.
Regulation of the Accumulation and Reduction of Nitrate by Nitrogen and Carbon Metabolites in Maize Seedlings
Authors: Sivasankar S. , Rothstein S. , Oaks A. 
Published: October 2016

Abstract
The accumulation and reduction of nitrate in the presence of the nitrogen metabolites asparagine (Asn) and glutamine (Gln) and the carbon metabolite sucrose (Suc) were examined in maize (Zea mays L.) seedlings in an attempt to separate their effects on the nitrate uptake system and the nitrate reduction system. After 8 h of exposure to nitrate in the presence of 1 mM Asn, tissue nitrate accumulation was reduced at 250 [mu]M external nitrate, but not at 5 mM Asn. The induction of nitrate reductase (NR) activity was reduced at both external nitrate concentrations. In the presence of 1 mM Gln or 1% Suc, tissue nitrate concentration was not significantly altered, but the induction of root NR activity was reduced or enhanced, respectively. The induction of root nitrite reductase (NiR) activity was also reduced in the presence of Asn or Gln and enhanced in the presence of Suc. Transcript levels of NR and NiR in roots were reduced in the presence of the amides and enhanced in the presence of Suc. When Suc was present in combination with either amide, there was complete relief from the inhibition of NiR transcription observed in the presence of amide alone. In the case of NR, however, this relief from inhibition was negligible. The inhibition of the induction of NR and NiR activities in the presence of Gln and Asn is a direct effect and is not the result of altered nitrate uptake in the presence of these metabolites.
ATP Hydrolysis Activity and Polymerization State of Ribulose-1,5-Bisphosphate Carboxylase Oxygenase Activase (Do the Effects of Mg2+, K+, and Activase Concentrations Indicate a Functional Similarity to Actin?)
Authors: Lilley R. M. , Portis Jr A. R. 
Published: October 2016

Abstract
The ATPase activity and fluoresence of ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) activase were determined over a range of MgCl2, KCl, and activase concentrations. Both salts promoted ADP release from ATP and intrinsic fluorescence enhancement by adenosine 5[prime]-[[gamma]-thio] triphosphate, but Mg2+ was about 10 times more effective than K+. ATPase and fluorescence enhancement both increased from zero to saturation within the same Mg2+ and K+ concentration ranges. At saturating concentrations (5 mM Mg2+ and 22 mM K+), the specific activity of ATPase (turnover time, about 1 s) and specific intrinsic fluorescence enhancement were maximal and unaffected by activase concentration above 1 [mu]M activase; below 1 [mu]M activase, both decreased sharply. These responses are remarkably similar to the behavior of actin. Intrinsic fluorescence enhancement of Rubisco activase reflects the extent of polymerization, showing that the smaller oligomer or monomer present in low-salt and activase concentrations is inactive in ATP hydrolysis. However, quenching of 1-anilinonapthaline-8-sulfonate fluorescence revealed that ADP and adenosine 5[prime]-[[gamma]-thio] triphosphate bind equally well to activase at low- and high-salt concentrations. This is consistent with an actin-like mechanism requiring a dynamic equilibrium between monomer and oligomers for ATP hydrolysis. The specific activation rate of substrate-bound decarbamylated Rubisco decreased at activase concentrations below 1 [mu]M. This suggests that a large oligomeric form of activase, rather than a monomer, interacts with Rubisco when performing the release of bound ribulose-1,5-bisphosphate from the inactive enzyme.
Subcellular Localization of Secondary Lipid Metabolites Including Fragrance Volatiles in Carnation Petals
Authors: Hudak K. A. , Thompson J. E. 
Published: October 2016

Abstract
Pulse-chase labeling of carnation (Dianthus caryophyllus L. cv Improved White Sim) petals with [14C]acetate has provided evidence for a hydrophobic subcompartment of lipid-protein particles within the cytosol that resemble oil bodies, are formed by blebbing from membranes, and are enriched in lipid metabolites (including fragrance volatiles) derived from membrane fatty acids. Fractionation of the petals during pulse-chase labeling revealed that radiolabeled fatty acids appear first in microsomal membranes and subsequently in cytosolic lipid-protein particles, indicating that the particles originate from membranes. This interpretation is supported by the finding that the cytosolic lipid-protein particles contain phospholipid as well as the same fatty acids found in microsomal membranes. Radiolabeled polar lipid metabolites (methanol/water-soluble) were detectable in both in situ lipid-protein particles isolated from the cytosol and those generated in vitro from isolated radiolabeled microsomal membranes. The lipid-protein particles were also enriched in hexanal, trans-2-hexenal, 1-hexanol, 3-hexen-1-ol, and 2-hexanol, volatiles of carnation flower fragrance that are derived from membrane fatty acids through the lipoxygenase pathway. Therefore, secondary lipid metabolites, including components of fragrance, appear to be formed within membranes of petal tissue and are subsequently released from the membrane bilayers into the cytosol by blebbing of lipid-protein particles.
A Plant Chloroplast Glutamyl Proteinase
Authors: Laing W. A. , Christeller J. T. 
Published: October 2016

Abstract
A glutamyl proteinase was partially purified from Percoll gradient-purified spinach (Spinacia oleracea) chloroplast preparations and appeared to be predominantly localized in the chloroplast stroma. The enzyme degraded casein, but of the 11 synthetic endopeptidase substrates tested, only benzyloxycarbonyl-leucine-leucine-glutamic acid-β-napthylamide was hydrolyzed at measurable rates. In addition, the enzyme cleaved the oxidized β-chain of insulin after a glutamic acid residue. There was no evidence that native ribulose-1,5-bisphosphate carboxylase/oxygenase was cleaved by this proteinase. The apparent Km for benzyloxycarbonyl-leucine-leucine-glutamic acid-[beta]NA at the pH optimum of 8.0 was about 1 mM. Cl-ions were required for both activity and stability. Of the proteinase inhibitors covering all four classes of the endopeptidases, only 4-(2-aminoethyl)-benzenesulfonyl-fluoride HCl and L-1-chloro-3-[4-tosylamido]-4-phenyl-2-butanone significantly inhibited the proteinase. The partially purified enzyme had a molecular weight of about 350,000 to 380,000, based on size-exclusion chromatography. The enzyme has both similar and distinctive properties to those of the bacterial glutamyl proteinases. To our knowledge, this is the first description of a plant glutamyl proteinase found predominantly or exclusively in the chloroplast.
Turnover of Galactans and Other Cell Wall Polysaccharides during Development of Flax Plants
Authors: Gorshkova T. A. , Chemikosova S. B. , Lozovaya V. V. , Carpita N. C. 
Published: October 2016

Abstract
We investigated the synthesis and turnover of cell wall polysaccharides of the flax (Linum usitatissimum L.) plant during development of the phloem fibers. One-month-old flax plants were exposed to a 40-min pulse with 14CO2 followed by 8-h, 24-h, and 1-month periods of chase with ambient CO2, and radioactivity in cell wall sugars was determined in various plant parts. The relative radioactivity of glucose in noncellulosic polysaccharides was the highest compared with all other cell wall sugars immediately after the pulse and decreased substantially during the subsequent chase. The relative radioactivities of the other cell wall sugars changed with differing rates, indicating turnover of specific polysaccharides. Notably, after 1 month of chase there was a marked decrease in the proportional mass and total radioactivity in cell wall galactose, indicating a long-term turnover of the galactans enriched in the fiber-containing tissues. The ratio of radiolabeled xylose to arabinose also increased during the chase, indicating a turnover of arabinose-containing polymers and interconversion to xylose. The pattern of label redistribution differed between organs, indicating that the cell wall turnover processes are tissue- and cell-specific.
Dissecting the Diphenylene Iodonium-Sensitive NAD(P)H:Quinone Oxidoreductase of Zucchini Plasma Membrane
Authors: Trost P. , Foscarini S. , Preger V. , Bonora P. , Vitale L. , Pupillo P. 
Published: October 2016

Abstract
Quinone oxidoreductase activities dependent on pyridine nucleotides are associated with the plasma membrane (PM) in zucchini (Cucurbita pepo L.) hypocotyls. In the presence of NADPH, lipophilic ubiquinone homologs with up to three isoprenoid units were reduced by intact PM vesicles with a Km of 2 to 7 [mu]M. Affinities for both NADPH and NADH were similar (Km of 62 and 51 [mu]M, respectively). Two NAD(P)H:quinone oxidoreductase forms were identified. The first, labeled as peak I in gel-filtration experiments, behaves as an intrinsic membrane complex of about 300 kD, it slightly prefers NADH over NADPH, it is markedly sensitive to the inhibitor diphenylene iodonium, and it is active with lipophilic quinones. The second form (peak II) is an NADPH-preferring oxidoreductase of about 90 kD, weakly bound to the PM. Peak II is diphenylene iodonium-insensitive and resembles, in many properties, the soluble NAD(P)H:quinone oxidoreductase that is also present in the same tissue. Following purification of peak I, however, the latter gave rise to a quinone oxidoreductase of the soluble type (peak II), based on substrate and inhibitor specificities and chromatographic and electrophoretic evidence. It is proposed that a redox protein of the same class as the soluble NAD(P)H:quinone oxidoreductase (F. Sparla, G. Tedeschi, and P. Trost [1996] Plant Physiol. 112:249–258) is a component of the diphenylene iodonium-sensitive PM complex capable of reducing lipophilic quinones.
β-1,3-Glucanase Is Cryoprotective in Vitro and Is Accumulated in Leaves during Cold Acclimation
Authors: Hincha D. K. , Meins Jr F. , Schmitt J. M. 
Published: October 2016

Abstract
We have used isolated spinach (Spinacea oleracea L.) thylakoid membranes to investigate the possible cryoprotective properties of class I β-1,3-glucanase (1,3-β-D-glucan 3-glucanohydrolase; EC 3.2.1.39) and chitinase. Class I β-1,3-glucanase that was purified from tobacco (Nicotiana tabacum L.) protected thylakoids against freeze-thaw injury in our in vitro assays, whereas class I chitinase from tobacco had no effect under the same conditions. The β-1,3-glucanase acted by reducing the influx of solutes into the membrane vesicles during freezing and thereby reduced osmotic stress and vesicle rupture during thawing. Western blots probed with antibodies directed against tobacco class I β-1,3-glucanase showed that in spinach and cabbage (Brassica oleracea L.) leaves an isoform of 41 kD was accumulated during frost hardening under natural conditions.
Experimental Analysis of Tassel Development in the Maize Mutant Tassel Seed 6
Authors: Irish E. E. 
Published: October 2016

Abstract
The maize (Zea mays L.) mutation Tassel seed 6 (Ts6) disrupts both sex determination in the tassel and the pattern of branching in inflorescences. This results in the formation of supernumerary florets in tassels and ears and in the development of pistils in tassel florets where they are normally aborted. A developmental analysis indicated that extra florets in Ts6 inflorescences are most likely the result of delayed determinacy in spikelet meristems, which then initiate additional floret meristems rather than initiating floral organs as in wild type. I have used culturing experiments to assay whether delayed determinacy of Ts6 mutant tassels is reflected in an altered timing of specific determination events. Length of the tassel was used as a developmental marker. These experiments showed that although Ts6 tassels elongate much more slowly than wild type, both mutant and wild-type tassels gained the ability to form flowers with organs of normal morphology in culture at the same time. In situ hybridization patterns of expression of the maize gene Kn, which is normally expressed in shoot meristems and not in determinate lateral organs, confirmed that additional meristems, rather than lateral organs, are initiated by spikelet meristems in Ts6 tassels.
Involvement of Thylakoid Overenergization in Tentoxin-Induced Chlorosis in Nicotiana spp
Authors: Holland N. , Evron Y. , Jansen MAK. , Edelman M. , Pick U. 
Published: October 2016

Abstract
The purpose of this work was to clarify the mechanism of tentoxin-induced chlorosis in Nicotiana spp. seedlings. We found that chlorosis does not correlate with the inhibition of chloroplast ATP synthesis in vivo, since it occurs at tentoxin concentrations far higher than that required for the inhibition of photophosphorylation measured in the same seedlings. However, tentoxin-induced chlorosis does correlate with in vivo overenergization of thylakoids. We show that tentoxin induces overenergization in intact plants and isolated thylakoids, probably via multiple interactions with ATP synthase. Furthermore, gramicidin D, a protonophore that relieves overenergization, also relieves chlorosis. Two lines of evidence suggest that reactive oxygen species may be involved in the process of chlorosis: ascorbate, a quencher of oxygen radicals, significantly protects against chlorosis, whereas transgenic Nicotiana spp. mutants overexpressing chloroplast superoxide dismutase are partially resistant to tentoxin-induced chlorosis. It is proposed that chlorosis in developing seedlings results from overenergization of thylakoids, which leads to the generation of oxygen radicals.