bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enhanced Expression and Activation of the Alternative Oxidase during Infection of Arabidopsis withPseudomonas syringae pv tomato1
Authors: Simons Bert H. , Millenaar Frank F. , Mulder Lonneke , Van Loon Leendert C. , Lambers Hans 
Published: July 2002

Abstract
Cyanide-resistant (“alternative”) respiration was studied in Arabidopsis during incompatible and compatible infection with Pseudomonas syringae pv tomato DC3000. Total leaf respiration increased as the leaves became necrotic, as did the cyanide-resistant component that was sensitive to salicylhydroxamic acid. Infiltration of leaves with an avirulent strain rapidly induced alternative oxidase (AOX) mRNA, whereas the increase was delayed in the compatible combination. The increase in mRNA correlated with the increase in AOX protein. Increased expression was confined to the infected leaves, in contrast to the pathogenesis-related protein-1, which was induced systemically. Virtually all of the AOX protein was in the reduced (high-activity) form. Using transgenic NahG and mutantnpr1-1 and etr1-1 plants, we established that the rapid induction of the AOX was associated with necrosis and that ethylene, but not salicylic acid, was required for its induction. Increased pyruvate levels in the infected leaves suggested that increased substrate levels were respired through the alternative pathway; however, in the control leaves and the infected leaves, respiration was not inhibited by salicylhydroxamic acid alone. Increased respiration appeared to be associated primarily with symptom expression rather than resistance reactions.
Aldehyde Oxidase and Xanthine Dehydrogenase in aflacca Tomato Mutant with Deficient Abscisic Acid and Wilty Phenotype1
Authors: Sagi Moshe , Fluhr Robert , Lips S. Herman 
Published: July 2002

Abstract
The flacca tomato (Lycopersicon esculentum) mutant displays a wilty phenotype as a result of abscisic acid (ABA) deficiency. The Mo cofactor (MoCo)-containing aldehyde oxidases (AO; EC 1.2.3.1) are thought to play a role in the final oxidation step required for ABA biosynthesis. AO and related MoCo-containing enzymes xanthine dehydrogenase (XDH; EC 1.2.1.37) and nitrate reductase (EC 1.6.6.1) were examined in extracts of the flacca tomato genotype and of wild-type (WT) roots and shoots. The levels of MoCo were found to be similar in both genotypes. No significant XDH or AO (MoCo-containing hydroxylases) activities were detected inflacca leaves; however, the mutant exhibited considerable MoCo-containing hydroxylase activity in the roots, which contained notable amounts of ABA. Native western blots probed with an antibody to MoCo-containing hydroxylases revealed substantial, albeit reduced, levels of cross-reactive protein in the flaccamutant shoots and roots. The ABA xylem-loading rate was significantly lower than that in the WT, indicating that the flacca is also defective in ABA transport to the shoot. Significantly, in vitro sulfurylation with Na2S reactivated preexisting XDH and AO proteins in extracts from flacca, particularly from the shoots, and superinduced the basal-level activity in the WT extracts. The results indicate that in flacca,MoCo-sulfurylase activity is impaired in a tissue-dependent manner.
Kinetic Analysis of Phospholipase C from Catharanthus roseus Transformed Roots Using Different Assays1
Authors: Hernández-Sotomayor S.M. Teresa , De Los Santos-Briones César , Muñoz-Sánchez J. Armando , Loyola-Vargas Victor M. 
Published: July 2002

Abstract
The properties of phospholipase C (PLC) partially purified from Catharanthus roseustransformed roots were analyzed using substrate lipids dispersed in phospholipid vesicles, phospholipid-detergent mixed micelles, and phospholipid monolayers spread at an air-water interface. Using [33P]phosphatidylinositol 4,5-bisphosphate (PIP2) of high specific radioactivity, PLC activity was monitored directly by measuring the loss of radioactivity from monolayers as a result of the release of inositol phosphate and its subsequent dissolution on quenching in the subphase. PLC activity was markedly affected by the surface pressure of the monolayer, with reduced activity at extremes of initial pressure. The optimum surface pressure for PIP2 hydrolysis was 20 mN/m. Depletion of PLC from solution by incubation with sucrose-loaded PIP2vesicles followed by ultracentrifugation demonstrated stable attachment of PLC to the vesicles. A mixed micellar system was established to assay PLC activity using deoxycholate. Kinetic analyses were performed to determine whether PLC activity was dependent on both bulk PIP2 and PIP2 surface concentrations in the micelles. The interfacial Michaelis constant was calculated to be 0.0518 mol fraction, and the equilibrium dissociation constant of PLC for the lipid was 45.5 μm. These findings will add to our understanding of the mechanisms of regulation of plant PLC.
Glucose Polyester Biosynthesis. Purification and Characterization of a Glucose Acyltransferase
Authors: Li Alice X. , Eannetta Nancy , Ghangas Gurdev S. , Steffens John C. 
Published: July 2002

Abstract
Glandular trichomes of the wild tomato species Lycopersicon pennellii secrete 2,3,4-O-tri-acyl-glucose (-Glc), which contributes to insect resistance. A Glc acyltransferase catalyzes the formation of diacyl-Glc by disproportionating two equivalents of 1-O-acyl-β-Glc, a high-energy molecule formed by a UDP-Glc dependent reaction. The acyltransferase was purified 4,900-fold from L. pennellii leaves by polyethylene glycol fractionation, diethylaminoethyl chromatography, concanavalin A affinity chromatography, and chromatofocusing. The acyltransferase possesses an isoelectric point of 4.8, a relative molecular mass around 110 kD, and is composed of 34- and 24-kD polypeptides as a heterotetramer. The 34- and 24-kD proteins were partially sequenced. The purified enzyme catalyzes both the disproportionation of 1-O-acyl-β-Glcs to generate 1,2-di-O-acyl-β-Glc and anomeric acyl exchange between 1-O-acyl-β-Glc and Glc.
Phosphatidylinositol 4-Kinase Associated with Spinach Plasma Membranes. Isolation and Characterization of Two Distinct Forms
Authors: Westergren Tomas , Ekblad Lars , Jergil Bengt , Sommarin Marianne 
Published: July 2002

Abstract
Highly purified plasma membranes from spinach (Spinacia oleracea L.) leaves contained phosphatidylinositol (PtdIns) kinase activity that was firmly associated with the membrane. The enzyme was solubilized by detergent treatment (2% [w/v] Triton X-100) and purified by heparin-Sepharose and Q-Sepharose chromatography. Two enzymically active fractions, QI and QII, both exhibiting PtdIns 4-kinase activity, were resolved and purified 100- to 300-fold over the plasma membrane. QI and QII shared similar high apparentK m values for ATP (approximately 0.45 mm) and PtdIns (approximately 0.2 mm) and were insensitive to inhibition by adenosine. While Mg2+ was the preferred divalent cation, Mn2+ could partly substitute in the reaction catalyzed by the QII enzyme but not in that catalyzed by QI. Mn2+ acted synergistically with suboptimal Mg2+ concentrations to activate not only the QII enzyme, but also to some extent QI. Both enzymes were inhibited by millimolar concentrations of Ca2+ and micromolar concentrations of wortmannin. The apparent molecular mass for QI was 120 kD, which was determined by SDS-PAGE and western blotting using an antibody against a peptide unique for lipid kinases and the binding of3H-wortmannin, and for QII 65 kD as determined by immunodetection and renaturation of PtdIns kinase activity in the 65-kD region of polyacrylamide gels.
Directed Mutation of the Rubisco Large Subunit of Tobacco Influences Photorespiration and Growth
Authors: Whitney Spencer M. , von Caemmerer Susanne , Hudson Graham S. , Andrews T. John 
Published: July 2002

Abstract
The gene for the large subunit of Rubisco was specifically mutated by transforming the chloroplast genome of tobacco (Nicotiana tabacum). Codon 335 was altered to encode valine instead of leucine. The resulting mutant plants could not grow without atmospheric CO2 enrichment. In 0.3% (v/v) CO2, the mutant and wild-type plants produced similar amounts of Rubisco but the extent of carbamylation was nearly twice as great in the mutants. The mutant enzyme's substrate-saturated CO2-fixing rate and its ability to distinguish between CO2 and O2 as substrates were both reduced to 25% of the wild type's values. Estimates of these parameters obtained from kinetic assays with the purified mutant enzyme were the same as those inferred from measurements of photosynthetic gas exchange with leaves of mutant plants. The Michaelis constants for CO2, O2, and ribulose-1,5-bisphosphate were reduced and the mutation enhanced oxygenase activity at limiting O2concentrations. Consistent with the reduced CO2 fixation rate at saturating CO2, the mutant plants grew slower than the wild type but they eventually flowered and reproduced apparently normally. The mutation and its associated phenotype were inherited maternally. The chloroplast-transformation strategy surmounts previous obstacles to mutagenesis of higher-plant Rubisco and allows the consequences for leaf photosynthesis to be assessed.
Identification of a Hsp70 Recognition Domain within the Rubisco Small Subunit Transit Peptide
Authors: Ivey Robert A. , Subramanian Chitra , Bruce Barry D. 
Published: July 2002

Abstract
The interaction between SStp, the transit peptide of the precursor protein to the small subunit of Rubisco (prSSU) and two Hsp70 molecular chaperones, Escherichia coli DnaK and pea (Pisum sativum) CSS1, was investigated in detail. Two statistical analyses were developed and used to investigate and predict regions of SStp recognized by DnaK. Both algorithms suggested that DnaK would have high affinity for the N terminus of SStp, moderate affinity for the central region, and low affinity for the C terminus. Furthermore, both algorithms predicted this affinity pattern for &gt;75% of the transit peptides analyzed in the chloroplast transit peptide (CHLPEP) database. In vitro association between SStp and these Hsp70s was confirmed by three independent assays: limited trypsin resistance, ATPase stimulation, and native gel shift. Finally, synthetic peptides scanning the length of SStp and C-terminal deletion mutants of SStp were used to experimentally map the region of greatest DnaK affinity to the N terminus. CSS1 displayed a similar affinity for the N terminus of SStp. The major stromal Hsp70s affinity for the N terminus of SStp and other transit peptides supports a molecular motor model in which the chaperone functions as an ATP-dependent translocase, committing chloroplast precursor proteins to unidirectional movement across the envelope.
The BtpA Protein Stabilizes the Reaction Center Proteins of Photosystem I in the Cyanobacterium Synechocystis sp. PCC 6803 at Low Temperature
Authors: Zak Elena , Pakrasi Himadri B. 
Published: July 2002

Abstract
Specific inhibition of photosystem I (PSI) was observed under low-temperature conditions in the cyanobacteriumSynechocystis sp. strain PCC 6803. Growth at 20°C caused inhibition of PSI activity and increased degradation of the PSI reaction center proteins PsaA and PsaB, while no significant changes were found in the level and activity of photosystem II (PSII). BtpA, a recently identified extrinsic thylakoid membrane protein, was found to be a necessary regulatory factor for stabilization of the PsaA and PsaB proteins under such low-temperature conditions. At normal growth temperature (30°C), the BtpA protein was present in the cell, and its genetic deletion caused an increase in the degradation of the PSI reaction center proteins. However, growth ofSynechocystis cells at 20°C or shifting of cultures grown at 30°C to 20°C led to a rapid accumulation of the BtpA protein, presumably to stabilize the PSI complex, by lowering the rates of degradation of the PsaA and PsaB proteins. A btpAdeletion mutant strain could not grow photoautotrophically at low temperature, and exhibited rapid degradation of the PSI complex after transfer of the cells from normal to low temperature.
Distinct Abscisic Acid Signaling Pathways for Modulation of Guard Cell versus Mesophyll Cell Potassium Channels Revealed by Expression Studies in Xenopus laevisOocytes
Authors: Sutton Fedora , Paul Sunil S. , Wang Xi-Qing , Assmann Sarah M. 
Published: July 2002

Abstract
Regulation of guard cell ion transport by abscisic acid (ABA) and in particular ABA inhibition of a guard cell inward K+current (I Kin) is well documented. However, little is known concerning ABA effects on ion transport in other plant cell types. Here we applied patch clamp techniques to mesophyll cell protoplasts of fava bean (Vicia faba cv Long Pod) plants and demonstrated ABA inhibition of an outward K+ current (I Kout). When mesophyll cell protoplast mRNA (mesophyll mRNA) was expressed in Xenopus laevisoocytes, I Kout was generated that displayed similar properties to I Kout observed from direct analysis of mesophyll cell protoplasts.I Kout expressed by mesophyll mRNA-injected oocytes was inhibited by ABA, indicating that the ABA signal transduction pathway observed in mesophyll cells was preserved in the frog oocytes. Co-injection of oocytes with guard cell protoplast mRNA and cRNA for KAT1, an inward K+ channel expressed in guard cells, resulted in I Kin that was similarly inhibited by ABA. However, oocytes co-injected with mesophyll mRNA and KAT1 cRNA produced I Kin that was not inhibited by ABA. These results demonstrate that the mesophyll-encoded signaling mechanism could not substitute for the guard cell pathway. These findings indicate that mesophyll cells and guard cells use distinct and different receptor types and/or signal transduction pathways in ABA regulation of K+ channels.
Effects of Elevated [CO2] and Nitrogen Nutrition on Cytokinins in the Xylem Sap and Leaves of Cotton
Authors: Yong Jean W.H. , Wong S. Chin , Letham D. Stuart , Hocart Charles H. , Farquhar Graham D. 
Published: July 2002

Abstract
We measured the level of xylem-derived cytokinins (CKs) entering a cotton leaf, and the CK levels in the same leaf, thus enabling xylem sap and foliar CKs to be compared concurrently. Although zeatin was the dominant CK in xylem sap, zeatin, dihydrozeatin, and N6-(2-isopentenyl) adenine were present in approximately equimolar levels in leaves. Elevated [CO2] (EC) has an effect on the levels of cytokinins in sap and leaf tissues. This effect was modulated by the two levels of root nitrogen nutrition (2 and 12 mm nitrate). Growth enhancement (70%) in EC over plants in ambient [CO2] (AC) was observed for both nitrogen nutrition treatments. Low-nitrogen leaves growing in EC exhibited photosynthetic acclimation, whereas there was no sign of photosynthetic acclimation in high-nitrogen grown leaves. Under these prevailing conditions, xylem sap and leaf tissues were obtained for CK analysis. Higher nitrogen nutrition increased the delivery per unit leaf area of CKs to the leaf at AC. EC caused a greater increase in CK delivery to the leaf at low nitrogen conditions (106%) than at high nitrogen conditions (17%). EC induced a significant increase in CK content in low-nitrogen leaves, whereas CK content in leaf tissues was similar for high-nitrogen leaves growing in AC and EC.
Dielectric Relaxation of Water and Water-Plasticized Biomolecules in Relation to Cellular Water Organization, Cytoplasmic Viscosity, and Desiccation Tolerance in Recalcitrant Seed Tissues
Authors: Sun Wendell Q. 
Published: July 2002

Abstract
To understand the relationship between the organization of cellular water, molecular interactions, and desiccation tolerance, dielectric behaviors of water and water-plasticized biomolecules in red oak (Quercus rubra) seeds were studied during dehydration. The thermally stimulated current study showed three dielectric dispersions: (a) the relaxation of loosely-bound water and small polar groups, (b) the relaxation of tightly-bound water, carbohydrate chains, large polar groups of macromolecules, and (c) the “freezing in” of molecular mobility (glassy state). Seven discrete hydration levels (water contents of 1.40, 0.55, 0.41, 0.31, 0.21, 0.13, and 0.08 g/g dry weight, corresponding to −1.5, −8, −11, −14, −24, −74, and −195 MPa, respectively) were identified according to the changes in thermodynamic and dielectric properties of water and water-plasticized biomolecules during dehydration. The implications of intracellular water organization for desiccation tolerance were discussed. Cytoplasmic viscosity increased exponentially at water content &lt; 0.40 g/g dry weight, which was correlated with the great relaxation slowdown of water-plasticized biomolecules, supporting a role for viscosity in metabolic shutdown during dehydration.
Pathogenesis of the Human Opportunistic PathogenPseudomonas aeruginosa PA14 in Arabidopsis
Authors: Plotnikova Julia M. , Rahme Laurence G. , Ausubel Frederick M. 
Published: July 2002

Abstract
The human opportunistic pathogen Pseudomonas aeruginosa strain PA14 is a multihost pathogen that can infect Arabidopsis. We found that PA14 pathogenesis in Arabidopsis involves the following steps: attachment to the leaf surface, congregation of bacteria at and invasion through stomata or wounds, colonization of intercellular spaces, and concomitant disruption of plant cell wall and membrane structures, basipetal movement along the vascular parenchyma, and maceration and rotting of the petiole and central bud. Distinctive features of P. aeruginosa pathogenesis are that the surface of mesophyll cell walls adopt an unusual convoluted or undulated appearance, that PA14 cells orient themselves perpendicularly to the outer surface of mesophyll cell walls, and that PA14 cells make circular perforations, approximately equal to the diameter of P. aeruginosa, in mesophyll cell walls. Taken together, our data show that P. aeruginosa strain PA14 is a facultative pathogen of Arabidopsis that is capable of causing local and systemic infection, which can result in the death of the infected plant.
Identification of Mutator Insertional Mutants of Starch-Branching Enzyme 2a in Corn
Authors: Blauth Susan L. , Yao Yuan , Klucinec Jeffery D. , Shannon Jack C. , Thompson Donald B. , Guiltinan Mark J. 
Published: July 2002

Abstract
Starch-branching enzymes (SBE) break the α-1,4 linkage of starch, re-attaching the chain to a glucan chain by an α-1,6 bond, altering starch structure. SBEs also facilitate starch accumulation by increasing the number of non-reducing ends on the growing chain. In maize (Zea mays), three isoforms of SBE have been identified. To examine the function of the SBEIIa isoform, a reverse genetics polymerase chain reaction-based screen was used to identify a mutant line segregating for a Mutator transposon withinSbe2a. To locate the insertion within the second exon ofSbe2a, the genomic sequence of Sbe2acontaining the promoter and 5′ end was isolated and sequenced. Plants homozygous for sbe2a::Mu have undetectable levels of Sbe2a transcripts and SBEIIa in their leaves. Characterization of leaf starch from sbe2a::Mumutants shows reduced branching similar to yet more extreme than that seen in kernels lacking SBEIIb activity. Characterization of endosperm starch from sbe2a::Mu mutants shows branching that is indistinguishable from wild-type controls. These mutant plants have a visible phenotype resembling accelerated senescence, which was correlated with the Mutator insertion withinSbe2a. This correlation suggests a specific role for SBEIIa in leaves, which may be necessary for normal plant development.
Regulation of psbA and psaE Expression by Light Quality in Synechocystis Species PCC 6803. A Redox Control Mechanism
Authors: Bissati Kamal El , Kirilovsky Diana 
Published: July 2002

Abstract
We investigated the influence of light of different wavelengths on the expression of the psbA gene, which encodes the D1 protein of the photosystem II and the psaE gene, which encodes the subunit Psa-E of the photosystem I, inSynechocystis sp PCC 6803. In an attempt to differentiate between a light-sensory and a redox-sensory signaling processes, the effect of orange, blue, and far-red light was studied in the wild-type and in a phycobilisome-less mutant. Transferring wild-type cells from one type of illumination to another induced changes in the redox state of the electron transport chain and inpsbA and psaE expression. Blue and far-red lights (which are preferentially absorbed by the photosystem I) induced an accumulation of psbA transcripts and a decrease of the psaE mRNA level. In contrast, orange light (which is preferentially absorbed by the photosystem II) induced a large accumulation of psaE transcripts and a decrease of psbA mRNA level. Transferring mutant cells from blue to orange light (or vice versa) had no effect either on the redox state of the electron transport chain or on the levels of psbAand psaE mRNAs. Thus, light quality seems to regulate expression of these genes via a redox sensory mechanism inSynechocystis sp PCC 6803 cells. Our data suggest that the redox state of one of the electron carriers between the plastoquinone pool and the photosystem I has opposite influences onpsbA and psaE expression. Its reduction induces accumulation of psaE transcripts, and its oxidation induces accumulation of psbA mRNAs.
Proteomic Analysis of Arabidopsis Seed Germination and Priming
Authors: Gallardo Karine , Job Claudette , Groot Steven P.C. , Puype Magda , Demol Hans , Vandekerckhove Joël , Job Dominique 
Published: July 2002

AbstractTo better understand seed germination, a complex developmental process, we developed a proteome analysis of the model plant Arabidopsis for which complete genome sequence is now available. Among about 1,300 total seed proteins resolved in two-dimensional gels, changes in the abundance (up- and down-regulation) of 74 proteins were observed during germination sensu stricto (i.e. prior to radicle emergence) and the radicle protrusion step. This approach was also used to analyze protein changes occurring during industrial seed pretreatments such as priming that accelerate seed germination and improve seedling uniformity. Several proteins were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry. Some of them had previously been shown to play a role during germination and/or priming in several plant species, a finding that underlines the usefulness of using Arabidopsis as a model system for molecular analysis of seed quality. Furthermore, the present study, carried out at the protein level, validates previous results obtained at the level of gene expression (e.g. from quantitation of differentially expressed mRNAs or analyses of promoter/reporter constructs). Finally, this approach revealed new proteins associated with the different phases of seed germination and priming. Some of them are involved either in the imbibition process of the seeds (such as an actin isoform or a WD-40 repeat protein) or in the seed dehydration process (e.g. cytosolic glyceraldehyde-3-phosphate dehydrogenase). These facts highlight the power of proteomics to unravel specific features of complex developmental processes such as germination and to detect protein markers that can be used to characterize seed vigor of commercial seed lots and to develop and monitor priming treatments.