bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differential Effect of Purified Spruce Chitinases and β-1,3-Glucanases on the Activity of Elicitors from Ectomycorrhizal Fungi
Authors: Salzer P. , Hubner B. , Sirrenberg A. , Hager A. 
Published: July 2002

Abstract
Two chitinases (EC 3.2.1.14) and two β-1,3-glucanases (EC 3.2.1.39) were purified from the culture medium of spruce (Picea abies [L.] Karst.) cells to study their role in modifying elicitors, cell walls, growth, and hyphal morphology of ectomycorrhizal fungi. The 36-kD class I chitinase (isoelectric point [pl] 8.0) and the 28-kD chitinase (pl 8.7) decreased the activity of elicitor preparations from Hebeloma crustuliniforme (Bull. ex Fries.) Quel., Amanita muscaria (L.) Pers., and Suillus variegatus (Sw.: Fr.) O.K., as demonstrated by using the elicitor-induced extracellular alkalinization in spruce cells as a test system. In addition, chitinases released monomeric products from the walls of these ectomycorrhizal fungi. The β-1,3-glucanases (35 kD, pl 3.7 and 3.9), in contrast, had little influence on the activity of the fungal elicitors and released only from walls of A. muscaria some polymeric products. Furthermore, chitinases alone and in combination with β-1,3-glucanases had no effect on the growth and morphology of the hyphae. Thus, it is suggested that apoplastic chitinases in the root cortex destroy elicitors from the ectomycorrhizal fungi without damaging the fungus. By this mechanism the host plant could attenuate the elicitor signal and adjust its own defense reactions to a level allowing symbiotic interaction.
Actin Filaments of Guard Cells Are Reorganized in Response to Light and Abscisic Acid
Authors: Eun S. O. , Lee Y. 
Published: July 2002

Abstract
We recently showed that treatment with actin antagonists perturbed stomatal behavior in Commelina communis L. leaf epidermis and therefore suggested that dynamic changes in actin are necessary for signal responses in guard cells (M. Kim, P.K. Hepler, S.O. Eun, K.-S. Ha, Y. Lee [1995] Plant Physiol 109: 1077–1084). Here we show that actin filaments of guard cells, visualized by immunofluorescence microscopy, change their distribution in response to physiological stimuli. When stomata were open under white-light illumination, actin filaments were localized in the cortex of guard cells, arranged in a pattern that radiates from the stomatal pore. In marked contrast, for guard cells of stomata closed by darkness or by abscisic acid, the actin organization was characterized by short fragments randomly oriented and diffusely labeled along the pore site. Upon abscisic acid treatment, the radial pattern of actin arrays in the illuminated guard cells began to disintegrate within a few minutes and was completely disintegrated in the majority of labeled guard cells by 60 min. Unlike actin filaments, microtubules of guard cells retained an unaltered organization under all conditions tested. These results further support the involvement of actin filaments in signal transduction pathways of guard cells.
Purification of the Plasma Membrane Ca2+-ATPase from Radish Seedlings by Calmodulin-Agarose Affinity Chromatography1
Authors: Bonza Cristina , Carnelli Antonella , De Michelis Maria Ida , Rasi-Caldogno Franca 
Published: July 2002

Abstract
The Ca2+-ATPase of the plasma membrane (PM) of germinating radish (Raphanus sativus L.) seeds was purified by calmodulin (CaM)-affinity chromatography using a batch procedure. PM purified by aqueous two-phase partitioning was solubilized withn-dodecyl β-d-maltoside and applied to a CaM-agarose matrix. After various washings with decreasing Ca2+ concentrations, the Ca2+-ATPase was eluted with 5 mm ethylenediaminetetraacetate (EDTA). The EDTA-eluted fraction contained about 25% of the loaded Ca2+-ATPase activity, with a specific activity 70-fold higher than that of the starting PM fraction. The EDTA-eluted fraction was highly enriched in a 133-kD polypeptide, which was identified as the PM Ca2+-ATPase by 125I-CaM overlay and fluorescein-isothiocyanate labeling. The PM Ca2+-ATPase cross-reacted with an antiserum against a putative Ca2+-ATPase of the Arabidopsis thalianachloroplast envelope.
Biological Activity of Reducing-End-Derivatized Oligogalacturonides in Tobacco Tissue Cultures1
Authors: Spiro Mark D. , Ridley Brent L. , Eberhard Stefan , Kates Keith A. , Mathieu Yves , O'Neill Malcolm A. , Mohnen Debra , Guern Jean , Darvill Alan , Albersheim Peter 
Published: July 2002

Abstract
The biological activity of reducing-end-modified oligogalacturonides was quantified in four tobacco (Nicotiana tabacum) tissue culture bioassays. The derivatives used were oligogalacturonides with the C-1 of their reducing end (a) covalently linked to a biotin hydrazide, (b) covalently linked to tyramine, (c) chemically reduced to a primary alcohol, or (d) enzymatically oxidized to a carboxylic acid. These derivatives were tested for their ability to (a) alter morphogenesis of N. tabacum cv Samsun thin cell-layer explants, (b) elicit extracellular alkalinization by suspension-cultured cv Samsun cells, (c) elicit extracellular alkalinization by suspension-cultured N. tabacum cv Xanthi cells, and (d) elicit H2O2 accumulation in the cv Xanthi cells. In all four bioassays, each of the derivatives had reduced biological activity compared with the corresponding underivatized oligogalacturonides, demonstrating that the reducing end is a key element for the recognition of oligogalacturonides in these systems. However, the degree of reduction in biological activity depends on the tissue culture system used and on the nature of the specific reducing-end modification. These results suggest that oligogalacturonides are perceived differently in each tissue culture system.
Cyst(e)ine Is the Transport Metabolite of Assimilated Sulfur from Bundle-Sheath to Mesophyll Cells in Maize Leaves1
Authors: Burgener Marta , Suter Marianne , Jones Stephanie , Brunold Christian 
Published: July 2002

Abstract
The intercellular distribution of the enzymes and metabolites of assimilatory sulfate reduction and glutathione synthesis was analyzed in maize (Zea mays L. cv LG 9) leaves. Mesophyll cells and strands of bundle-sheath cells from second leaves of 11-d-old maize seedlings were obtained by two different mechanical-isolation methods. Cross-contamination of cell preparations was determined using ribulose bisphosphate carboxylase (EC 4.1.1.39) and nitrate reductase (EC 1.6.6.1) as marker enzymes for bundle-sheath and mesophyll cells, respectively. ATP sulfurylase (EC 2.7.7.4) and adenosine 5′-phosphosulfate sulfotransferase activities were detected almost exclusively in the bundle-sheath cells, whereas GSH synthetase (EC 6.3.2.3) and cyst(e)ine, γ-glutamylcysteine, and glutathione were located predominantly in the mesophyll cells. Feeding experiments using [35S]sulfate with intact leaves indicated that cyst(e)ine was the transport metabolite of reduced sulfur from bundle-sheath to mesophyll cells. This result was corroborated by tracer experiments, which showed that isolated bundle-sheath strands fed with [35S]sulfate secreted radioactive cyst(e)ine as the sole thiol into the resuspending medium. The results presented in this paper show that assimilatory sulfate reduction is restricted to the bundle-sheath cells, whereas the formation of glutathione takes place predominantly in the mesophyll cells, with cyst(e)ine functioning as a transport metabolite between the two cell types.
Osmotic Water Permeability of Isolated Protoplasts. Modifications during Development1
Authors: Ramahaleo Tiana , Morillon Raphaël , Alexandre Joël , Lassalles Jean-Paul 
Published: July 2002

Abstract
A transference chamber was developed to measure the osmotic water permeability coefficient (Pos) in protoplasts 40 to 120 μm in diameter. The protoplast was held by a micropipette and submitted to a steep osmotic gradient created in the transference chamber.Pos was derived from the changes in protoplast dimensions, as measured using a light microscope. Permeabilities were in the range 1 to 1000 μm s−1 for the various types of protoplasts tested. The precision forPos was ≤40%, and within this limit, no asymmetry in the water fluxes was observed. Measurements on protoplasts isolated from 2- to 5-d-old roots revealed a dramatic increase inPos during root development. A shift inPos from 10 to 500 μm s−1occurred within less than 48 h. This phenomenon was found in maize (Zea mays), wheat (Triticum aestivum), and rape (Brassica napus) roots. These results show that early developmental processes modify water-transport properties of the plasma membrane, and that the transference chamber is adapted to the study of water-transport mechanisms in native membranes.
Acclimation of Arabidopsis Leaves Developing at Low Temperatures. Increasing Cytoplasmic Volume Accompanies Increased Activities of Enzymes in the Calvin Cycle and in the Sucrose-Biosynthesis Pathway1
Authors: Strand Åsa , Hurry Vaughan , Henkes Stefan , Huner Norman , Gustafsson Petter , Gardeström Per , Stitt Mark 
Published: July 2002

Abstract
Photosynthetic and metabolic acclimation to low growth temperatures were studied in Arabidopsis (Heynh.). Plants were grown at 23°C and then shifted to 5°C. We compared the leaves shifted to 5°C for 10 d and the new leaves developed at 5°C with the control leaves on plants that had been left at 23°C. Leaf development at 5°C resulted in the recovery of photosynthesis to rates comparable with those achieved by control leaves at 23°C. There was a shift in the partitioning of carbon from starch and toward sucrose (Suc) in leaves that developed at 5°C. The recovery of photosynthetic capacity and the redirection of carbon to Suc in these leaves were associated with coordinated increases in the activity of several Calvin-cycle enzymes, even larger increases in the activity of key enzymes for Suc biosynthesis, and an increase in the phosphate available for metabolism. Development of leaves at 5°C also led to an increase in cytoplasmic volume and a decrease in vacuolar volume, which may provide an important mechanism for increasing the enzymes and metabolites in cold-acclimated leaves. Understanding the mechanisms underlying such structural changes during leaf development in the cold could result in novel approaches to increasing plant yield.
white anther: A Petunia Mutant That Abolishes Pollen Flavonol Accumulation, Induces Male Sterility, and Is Complemented by a Chalcone Synthase Transgene1
Authors: Napoli Carolyn A. , Fahy Deirdre , Wang Huai-Yu , Taylor Loverine P. 
Published: July 2002

Abstract
A mutation in an inbred line of petunia (Petunia hybrida) produces a reduction in the deep-purple corolla pigmentation and changes the anther color from yellow to white. In addition, the mutant, designated white anther (wha), is functionally male sterile. The inability of pollen fromwha plants to germinate in vitro provides a physiological basis for the lack of seed set observed in self-crosses of the mutant. Biochemical complementation with nanomolar amounts of kaempferol, a flavonol aglycone, confirms that the inability of thewha pollen to germinate is due to a lack of this essential compound. Transgenic complementation with a functionalChsA (ChalconesynthaseA) cDNA suggests that the genetic lesion responsible for the wha phenotype is in Chs, the gene for the first enzyme in the flavonol biosynthesis pathway. The genetic background of the parental line, as well as the pollen phenotype, allowed us to deduce that thewha mutation is in ChsA. To our knowledge, wha is the first induced, nontransgenicChs mutant described in petunia, and analysis of the mutation confirms earlier molecular and genetic observations that only two Chs genes (A and J) are expressed in reproductive tissues and that they are differentially regulated in corolla and anther.
Purification and Characterization of the Reconstitutively Active Citrate Carrier from Maize Mitochondria1
Authors: Genchi Giuseppe , Spagnoletta Anna , De Santis Aurelio , Stefanizzi Luisa , Palmieri Ferdinando 
Published: July 2002

Abstract
The citrate carrier from maize(Zea mays) shoot mitochondria was solubilized with Triton X-100 and purified by sequential chromatography on hydroxyapatite and hydroxyapatite/celite in the presence of cardiolipin. SDS-gel electrophoresis of the purified fraction showed a single polypeptide band with an apparent molecular mass of 31 kD. When reconstituted into liposomes, the citrate carrier catalyzed a pyridoxal 5′-P-sensitive citrate/citrate exchange. It was purified 224-fold with a recovery of 50% and a protein yield of 0.22% with respect to the mitochondrial extract. In the reconstituted system the purified citrate carrier catalyzed a first-order reaction of citrate/citrate (0.065 min−1) or citrate/malate exchange (0.075 min−1). Among the various substrates and inhibitors tested, the reconstituted protein transported citrate,cis-aconitate, isocitrate, l-malate, succinate, malonate, glutarate, α-ketoglutarate, oxaloacetate, and α-ketoadipate and was inhibited by pyridoxal 5′-P, phenylisothiocyanate, mersalyl, andp-hydroxymercuribenzoate (but notN-ethylmaleimide), 1,2,3-benzentricarboxylate, benzylmalonate, and butylmalonate. The activation energy of the citrate/citrate exchange was 66.5 kJ/mol between 10°C and 35°C; the half-saturation constant (Km) for citrate was 0.65 ± 0.05 mm and the maximal rate (Vmax) of the citrate/citrate exchange was 13.0 ± 1.0 μmol min−1 mg−1 protein at 25°C.
Modulation of Rubisco Activity during the Diurnal Phases of the Crassulacean Acid Metabolism Plant Kalanchoëdaigremontiana
Authors: Maxwell Kate , Borland Anne M. , Haslam Richard P. , Helliker Brent R. , Roberts Andrew , Griffiths Howard 
Published: July 2002

Abstract
The regulation of Rubisco activity was investigated under high, constant photosynthetic photon flux density during the diurnal phases of Crassulacean acid metabolism in Kalanchoëdaigremontiana Hamet et Perr. During phase I, a significant period of nocturnal, C4-mediated CO2 fixation was observed, with the generated malic acid being decarboxylated the following day (phase III). Two periods of daytime atmospheric CO2 fixation occurred at the beginning (phase II, C4–C3 carboxylation) and end (phase IV, C3–C4 carboxylation) of the day. During the 1st h of the photoperiod, when phosphoenolpyruvate carboxylase was still active, the highest rates of atmospheric CO2 uptake were observed, coincident with the lowest rates of electron transport and minimal Rubisco activity. Over the next 1 to 2 h of phase II, carbamylation increased rapidly during an initial period of decarboxylation. Maximal carbamylation (70%–80%) was reached 2 h into phase III and was maintained under conditions of elevated CO2 resulting from malic acid decarboxylation. Initial and total Rubisco activity increased throughout phase III, with maximal activity achieved 9 h into the photoperiod at the beginning of phase IV, as atmospheric CO2 uptake recommenced. We suggest that the increased enzyme activity supports assimilation under CO2-limited conditions at the start of phase IV. The data indicate that Rubisco activity is modulated in-line with intracellular CO2 supply during the daytime phases of Crassulacean acid metabolism.
The Multiple Roles of Conserved Arginine 286 of 1-Aminocyclopropane-1-Carboxylate Synthase. Coenzyme Binding, Substrate Binding, and Beyond
Authors: Zhou Huiqing , Wang Hong Wei , Zhu Ken , Sui Sen Fang , Xu Peilin , Yang Shang Fa , Li Ning 
Published: July 2002

Abstract
A pyridoxal 5′-phosphate (PLP)-dependent enzyme, 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (S-adenosyl-l-Met methylthioadenosine-lyase, EC 4.4.1.14), catalyzes the conversion ofS-adenosyl-l-methionine (AdoMet) to ACC. A tomato ACC synthase isozyme (LE-ACS2) with a deletion of 46 amino acids at the C terminus was chosen as the control enzyme for the study of the function of R286 in ACC synthase. R286 of the tomato ACC synthase was mutated to a leucine via site-directed mutagenesis. The ACC synthase mutant R286L was purified using a simplified two-step purification protocol. Circular dichroism (CD) analysis indicated that the overall three-dimensional structure of the mutant was indistinguishable from that of the control enzyme. Fluorescence spectroscopy revealed that the binding affinity of R286L ACC synthase for its cofactor PLP was reduced 20- to 25-fold compared with control. Kinetic analysis of R286L showed that this mutant ACC synthase had a significantly reduced turnover number (k cat) of 8.2 × 10−3 s−1 and an increasedK m of 730 μm for AdoMet, leading to an 8,000-fold decrease in overall catalytic efficiency compared with the control enzyme. Thus, R286 of tomato ACC synthase is involved in binding both PLP and AdoMet.
Reduction and Coordination of Arsenic in Indian Mustard
Authors: Pickering Ingrid J. , Prince Roger C. , George Martin J. , Smith Robert D. , George Graham N. , Salt David E. 
Published: July 2002

Abstract
The bioaccumulation of arsenic by plants may provide a means of removing this element from contaminated soils and waters. However, to optimize this process it is important to understand the biological mechanisms involved. Using a combination of techniques, including x-ray absorption spectroscopy, we have established the biochemical fate of arsenic taken up by Indian mustard (Brassica juncea). After arsenate uptake by the roots, possibly via the phosphate transport mechanism, a small fraction is exported to the shoot via the xylem as the oxyanions arsenate and arsenite. Once in the shoot, the arsenic is stored as an AsIII-tris-thiolate complex. The majority of the arsenic remains in the roots as an AsIII-tris-thiolate complex, which is indistinguishable from that found in the shoots and from AsIII-tris-glutathione. The thiolate donors are thus probably either glutathione or phytochelatins. The addition of the dithiol arsenic chelator dimercaptosuccinate to the hydroponic culture medium caused a 5-fold-increased arsenic level in the leaves, although the total arsenic accumulation was only marginally increased. This suggests that the addition of dimercaptosuccinate to arsenic-contaminated soils may provide a way to promote arsenic bioaccumulation in plant shoots, a process that will be essential for the development of an efficient phytoremediation strategy for this element.
The Role of Microtubules in Guard Cell Function
Authors: Marcus Adam I. , Moore Richard C. , Cyr Richard J. 
Published: July 2002

Abstract
Guard cells are able to sense a multitude of environmental signals and appropriately adjust the stomatal pore to regulate gas exchange in and out of the leaf. The role of the microtubule cytoskeleton during these stomatal movements has been debated. To help resolve this debate, in vivo stomatal aperture assays with different microtubule inhibitors were performed. We observed that guard cells expressing the microtubule-binding green fluorescent fusion protein (green fluorescent protein::microtubule binding domain) fail to open for all major environmental triggers of stomatal opening. Furthermore, guard cells treated with the anti-microtubule drugs, propyzamide, oryzalin, and trifluralin also failed to open under the same environmental conditions. The inhibitory conditions caused by green fluorescent protein::microtubule binding domain and these anti-microtubule drugs could be reversed using the proton pump activator, fusicoccin. Therefore, we conclude that microtubules are involved in an upstream event prior to the ionic fluxes leading to stomatal opening. In a mechanistic manner, evidence is presented to implicate a microtubule-associated protein in this putative microtubule-based signal transduction event.
Overexpression of Acyl Carrier Protein-1 Alters Fatty Acid Composition of Leaf Tissue in Arabidopsis
Authors: Branen Jill K. , Chiou Tzyy-Jen , Engeseth Nicki J. 
Published: July 2002

Abstract
Acyl carrier protein (ACP) is a small (9 kD) acidic protein that is an essential cofactor in plant fatty acid biosynthesis. Most plants have several isoforms of ACP, some of which are expressed constitutively and others that appear to be more tissue specific. Although the critical role of ACP in fatty acid biosynthesis has been established, the role of the diverse number of isoforms has yet to be elucidated. We have generated transgenic Arabidopsis plants that express high levels of ACP-1, a seed-predominant ACP isoform, in leaf tissue under control of the cauliflower mosaic virus 35S promoter. Western and northern analysis of these plants demonstrate 3- to 8-fold increased expression of this isoform in leaf tissue, but no significant changes in seed. Analysis of the fatty acid composition of leaf tissue revealed that overexpression of ACP-1 in leaf tissue alters fatty acid composition. Significant decreases in levels of 16:3 were noted along with increases in 18:3. These findings represent the first in vivo report that overexpression of an ACP isoform results in changes in fatty acid composition in plants.
Solvation energetics and conformational change in EF‐hand proteins
Authors: Ababou Abdessamad , Desjarlais John R. 
Published: July 2002

AbstractCalmodulin and other members of the EF‐hand protein family are known to undergo major changes in conformation upon binding Ca2+. However, some EF‐hand proteins, such as calbindin D9k, bind Ca2+ without a significant change in conformation. Here, we show the importance of a precise balance of solvation energetics to conformational change, using mutational analysis of partially buried polar groups in the N‐terminal domain of calmodulin (N‐cam). Several variants were characterized using fluorescence, circular dichroism, and NMR spectroscopy. Strikingly, the replacement of polar side chains glutamine and lysine at positions 41 and 75 with nonpolar side chains leads to dramatic enhancement of the stability of the Ca2+‐free state, a corresponding decrease in Ca2+‐binding affinity, and an apparent loss of ability to change conformation to the open form. The results suggest a paradigm for conformational change in which energetic strain is accumulated in one state in order to modulate the energetics of change to the alternative state.
Role for cysteine residues in the in vivo folding and assembly of the phage P22 tailspike
Authors: Haase‐Pettingell Cameron , Betts Scott , Raso Stephen W. , Stuart Lisa , Robinson Anne , King Jonathan 
Published: July 2002

AbstractThe predominantly β‐sheet phage P22 tailspike adhesin contains eight reduced cysteines per 666 residue chain, which are buried and unreactive in the native trimer. In the pathway to the native trimer, both in vivo and in vitro transient interchain disulfide bonds are formed and reduced. This occurs in the protrimer, an intermediate in the formation of the interdigitated β‐sheets of the trimeric tailspike. Each of the eight cysteines was replaced with serine by site‐specific mutagenesis of the cloned P22 tailspike gene and the mutant genes expressed in Escherichia coli. Although the yields of native‐like Cys>Ser proteins varied, sufficient soluble trimeric forms of each of the eight mutants accumulated to permit purification. All eight single Cys>Ser mature proteins maintained the high thermostability of the wild type, as well as the wild‐type biological activity in forming infectious virions. Thus, these cysteine thiols are not required for the stability or activity of the native state. When their in vivo folding and assembly kinetics were examined, six of the mutant substitutions—C267S, C287S, C458S, C613S, and C635S—were significantly impaired at higher temperatures. Four—C290S, C496, C613S, and C635—showed significantly impaired kinetics even at lower temperatures. The in vivo folding of the C613S/C635S double mutant was severely defective independent of temperature. Since the trimeric states of the single Cys>Ser substituted chains were as stable and active as wild type, the impairment of tailspike maturation presumably reflects problems in the in vivo folding or assembly pathways. The formation or reduction of the transient interchain disulfide bonds in the protrimer may be the locus of these kinetic functions.