scholarly journals Identification and characterization of plant glycerophosphodiester phosphodiesterase

2004 ◽  
Vol 379 (3) ◽  
pp. 601-607 ◽  
Author(s):  
Benoît van der REST ◽  
Norbert ROLLAND ◽  
Anne-Marie BOISSON ◽  
Myriam FERRO ◽  
Richard BLIGNY ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Cell Walls ◽  
Expression System ◽  
Gel Filtration ◽  
Cell Suspension Cultures ◽  
Cell Wall Proteins ◽  
Ammonium Sulphate Precipitation ◽  
Carrot Cell ◽  
Glycerophosphodiester Phosphodiesterase

GPX-PDE (glycerophosphodiester phosphodiesterase; EC 3.1.4.46) is a relatively poorly characterized enzyme that catalyses the hydrolysis of various glycerophosphodiesters (glycerophosphocholine, glycerophosphoethanolamine, glycerophosphoglycerol, glycerophosphoserine and bis-glycerophosphoglycerol), releasing sn-glycerol 3-phosphate and the corresponding alcohol. In a previous study, we demonstrated the existence of a novel GPX-PDE in the cell walls and vacuoles of plant cells. Since no GPX-PDE had been identified in any plant organism, the purification of GPX-PDE from carrot cell walls was attempted. After extraction of cell wall proteins from carrot cell suspension cultures with CaCl2, GPX-PDE was purified up to 2700-fold using, successively, ammonium sulphate precipitation, gel filtration and concanavalin A–Sepharose. Internal sequence analysis of a 55 kDa protein identified in the extract following 2700-fold purification revealed strong similarity to the primary sequence of GLPQ, a bacterial GPX-PDE. To confirm the identity of plant GPX-PDE, an Arabidopsis thaliana cDNA similar to that encoding the bacterial GPX-PDE was cloned and overexpressed in a bacterial expression system, and was used to raise antibodies against the putative Arabidopsis thaliana GPX-PDE. Immunochemical assays performed on carrot cell wall proteins extracted by CaCl2 treatment showed a strong correlation between GPX-PDE activity and detection of the 55 kDa protein, validating the identity of the plant GPX-PDE. Finally, various properties of the purified enzyme were investigated. GPX-PDE is a multimeric enzyme, specific for glycerophosphodiesters, exhibiting a Km of 36 µM for glycerophosphocholine and active within a wide pH range (from 4 to 10). Since these properties are similar to those of GLPQ, the bacterial GPX-PDE, the similarities between plant and bacterial enzymes are also discussed.

scholarly journals Changes in cell ultrastructure and morphology of Arabidopsis thaliana roots after coumarins treatment

2014 ◽  
Vol 70 (3) ◽  
pp. 187-198
Author(s):  
Ewa Kupidłowska
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Cell Walls ◽  
Root Elongation ◽  
Inhibitory Effect ◽  
Cell Ultrastructure ◽  
Radial Expansion ◽  
Elongation Zone ◽  
Cortical Cells ◽  
Mother Cell Wall

The ultrastructure and morphology of roots treated with coumarin and umbelliferone as well as the reversibility of the coumarins effects caused by exogenous GA, were studied in <em>Arabidopsis thaliana</em>. Both coumarins suppressed root elongation and appreciably stimulated radial expansion of epidermal and cortical cells in the upper part of the meristem and in the elongation zone. The gibberellic acid applied simultaneously with coumarins decreased their inhibitory effect on root elongation and reduced cells swelling.Microscopic observation showed intensive vacuolization of cells and abnormalities in the structure of the Golgi stacks and the nuclear envelope. The detection of active acid phosphatase in the cytosol of swollen cells indicated increased membrane permeability. Significant abnormalities of newly formed cell walls, e.g. the discontinuity of cellulose layer, uncorrect position of walls and the lack of their bonds with the mother cell wall suggest that coumarins affected the cytoskeleton.

scholarly journals Structure and Biomechanics during Xylem Vessel Transdifferentiation in Arabidopsis thaliana

Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1715
Author(s):  
Eleftheria Roumeli ◽  
Leah Ginsberg ◽  
Robin McDonald ◽  
Giada Spigolon ◽  
Rodinde Hendrickx ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Cell Walls ◽  
Turgor Pressure ◽  
Plant Cells ◽  
Building Blocks ◽  
Xylem Vessel ◽  
Primary Cell Wall ◽  
Individual Plant ◽  
Vessel Elements

Individual plant cells are the building blocks for all plantae and artificially constructed plant biomaterials, like biocomposites. Secondary cell walls (SCWs) are a key component for mediating mechanical strength and stiffness in both living vascular plants and biocomposite materials. In this paper, we study the structure and biomechanics of cultured plant cells during the cellular developmental stages associated with SCW formation. We use a model culture system that induces transdifferentiation of Arabidopsis thaliana cells to xylem vessel elements, upon treatment with dexamethasone (DEX). We group the transdifferentiation process into three distinct stages, based on morphological observations of the cell walls. The first stage includes cells with only a primary cell wall (PCW), the second covers cells that have formed a SCW, and the third stage includes cells with a ruptured tonoplast and partially or fully degraded PCW. We adopt a multi-scale approach to study the mechanical properties of cells in these three stages. We perform large-scale indentations with a micro-compression system in three different osmotic conditions. Atomic force microscopy (AFM) nanoscale indentations in water allow us to isolate the cell wall response. We propose a spring-based model to deconvolve the competing stiffness contributions from turgor pressure, PCW, SCW and cytoplasm in the stiffness of differentiating cells. Prior to triggering differentiation, cells in hypotonic pressure conditions are significantly stiffer than cells in isotonic or hypertonic conditions, highlighting the dominant role of turgor pressure. Plasmolyzed cells with a SCW reach similar levels of stiffness as cells with maximum turgor pressure. The stiffness of the PCW in all of these conditions is lower than the stiffness of the fully-formed SCW. Our results provide the first experimental characterization of the mechanics of SCW formation at single cell level.

scholarly journals Purification and partial characterization of a peroxidase from plant cell cultures of Cassia didymobotrya and biotransformation studies1

1998 ◽  
Vol 331 (2) ◽  
pp. 513-519 ◽  
Author(s):  
Alberto VITALI ◽  
Bruno BOTTA ◽  
Giuliano DELLE MONACHE ◽  
Sabrina ZAPPITELLI ◽  
Paola RICCIARDI ◽  
...  
Keyword(s):  
Cell Wall ◽  
Culture Medium ◽  
Gel Filtration ◽  
Cell Suspension Cultures ◽  
High Specificity ◽  
Coniferyl Alcohol ◽  
Terminal Sequence ◽  
Sds Page ◽  
Plant Peroxidases

An acidic peroxidase (EC 1.11.1.7) produced by cell suspension cultures of Cassia didymobotrya(wild senna) was purified from culture medium collected on the 29th day. The enzyme was shown to be a glycoprotein with a pI of 3.5, a molecular mass of approx. 43 kDa by SDS/PAGE and 50 kDa by gel filtration. The N-terminal sequence was very similar to those of other plant peroxidases. The peroxidase was characterized by a high specificity towards coniferyl alcohol and other natural phenolics such as guaiacol and ferulic and caffeic acids. These findings suggest that the enzyme is involved in lignification processes of the cell wall. Moreover, the enzyme was able to catalyse the oxidation of 4,3´,4´-trihydroxychalcone and 4,3´,4´-trihydroxy-3-methoxychalcone to the corresponding 3,3´-biflavanones, as mixtures of racemic and mesoforms.

scholarly journals Cytokinesis in fra2 Arabidopsis thaliana p60-katanin Mutant: Defects in Cell Plate/Daughter Wall Formation

2021 ◽  
Author(s):  
Emmanuel Panteris ◽  
Anna Kouskouveli ◽  
Dimitris Pappas ◽  
Ioannis-Dimosthenis S. Adamakis
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Cell Walls ◽  
Higher Plants ◽  
Cell Plate ◽  
Wild Type ◽  
Microtubule Organization ◽  
Root Cells ◽  
Wall Formation ◽  
In The Wild

Cytokinesis is accomplished in higher plants by the phragmoplast, creating and conducting the cell plate, to separate daughter nuclei by a new cell wall. The microtubule-severing enzyme p60-katanin plays an important role in the centrifugal expansion and timely disappearance of phragmoplast microtubules. Consequently, aberrant structure and delayed expansion rate of the phragmoplast occur in p60-katanin mutants. Here, the consequences of p60-katanin malfunction in cell plate/daughter wall formation were investigated by transmission electron microscopy (TEM), while deviations in the chemical composition of cell plate/new cell wall were identified by immunolabeling and confocal microscopy, in root cells of the fra2 Arabidopsis thaliana mutant. It was found that, apart from defective phragmoplast microtubule organization, cell plates/new cell walls appeared also faulty in structure, being unevenly thick and perforated by large gaps. In addition, demethylesterified homogalacturonans were prematurely present in fra2 cell plates, while callose content was significantly lower than in the wild-type. Furthermore, KNOLLE syntaxin disappeared from newly formed cell walls in fra2 earlier than in the wild-type. Taken together, these observations indicate that delayed cytokinesis, due to faulty phragmoplast organization and expansion, results in a loss of synchronization between cell plate growth and its chemical maturation.

scholarly journals Post-Synthetic Reduction of Pectin Methylesterification Causes Morphological Abnormalities and Alterations to Stress Response in Arabidopsis thaliana

Plants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1558
Author(s):  
Nathan T. Reem ◽  
Lauran Chambers ◽  
Ning Zhang ◽  
Siti Farah Abdullah ◽  
Yintong Chen ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Plant Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Normal Growth ◽  
Pectin Methylesterase ◽  
Dwarf Phenotype ◽  
Response To Stress ◽  
Defense Response Genes

Pectin is a critical component of the plant cell wall, supporting wall biomechanics and contributing to cell wall signaling in response to stress. The plant cell carefully regulates pectin methylesterification with endogenous pectin methylesterases (PMEs) and their inhibitors (PMEIs) to promote growth and protect against pathogens. We expressed Aspergillus nidulans pectin methylesterase (AnPME) in Arabidopsis thaliana plants to determine the impacts of methylesterification status on pectin function. Plants expressing AnPME had a roughly 50% reduction in methylester content compared with control plants. AnPME plants displayed a severe dwarf phenotype, including small, bushy rosettes and shorter roots. This phenotype was caused by a reduction in cell elongation. Cell wall composition was altered in AnPME plants, with significantly more arabinose and significantly less galacturonic acid, suggesting that plants actively monitor and compensate for altered pectin content. Cell walls of AnPME plants were more readily degraded by polygalacturonase (PG) alone but were less susceptible to treatment with a mixture of PG and PME. AnPME plants were insensitive to osmotic stress, and their susceptibility to Botrytis cinerea was comparable to wild type plants despite their compromised cell walls. This is likely due to upregulated expression of defense response genes observed in AnPME plants. These results demonstrate the importance of pectin in both normal growth and development, and in response to biotic and abiotic stresses.

scholarly journals Di-isodityrosine, a novel tetrametric derivative of tyrosine in plant cell wall proteins: a new potential cross-link

1996 ◽  
Vol 315 (1) ◽  
pp. 323-327 ◽  
Author(s):  
Jeffrey D. BRADY ◽  
Ian H. SADLER ◽  
Stephen C. FRY
Keyword(s):  
Cell Culture ◽  
Cell Wall ◽  
Amino Acid ◽  
Plant Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Cell Wall Proteins ◽  
Cross Link ◽  
Nmr Spectrometry

A novel amino acid, di-isodityrosine, has been isolated from hydrolysates of cell walls of tomato cell culture. Analysis by UV spectrometry, partial derivatization with 2,4-dinitrofluorobenzene and mass and NMR spectrometry show that the compound is composed to two molecules of isodityrosine, joined by a biphenyl linkage. The possible reactions involved in the formation of this molecule in vivo are discussed, as is the possibility that it could form an interpolypeptide linkage between cell wall proteins such as extensin, and hence aid in the insolubilization of the protein in the wall.

scholarly journals Proteomics of loosely bound cell wall proteins ofArabidopsis thaliana cell suspension cultures: A critical analysis

2003 ◽  
Vol 24 (1920) ◽  
pp. 3421-3432 ◽  
Author(s):  
Gisèle Borderies ◽  
Elisabeth Jamet ◽  
Claude Lafitte ◽  
Michel Rossignol ◽  
Alain Jauneau ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Suspension ◽  
Critical Analysis ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Cell Wall Proteins ◽  
Ofarabidopsis Thaliana

Composition and properties of the cell wall of Methanospirillum hungatii

1980 ◽  
Vol 26 (2) ◽  
pp. 115-120 ◽  
Author(s):  
G. D. Sprott ◽  
R. C. McKellar
Keyword(s):  
Amino Acids ◽  
Cell Wall ◽  
Cell Walls ◽  
Weight Fraction ◽  
Dry Weight ◽  
Wall Material ◽  
Bond Breaking ◽  
High Molecular Weight Fraction ◽  
Cell Wall Proteins ◽  
Isolated Cell

Dithiothreitol reacted, at pH 9.0, with the isolated cell walls of Methanospirillum hungatii, to release about 23% of the cell wall dry weight as a high molecular weight fraction (> 0.5 million daltons). Untreated walls consisted of 70% amino acids, 11% lipid, and 6.6% carbohydrate. Sugars were identified as rhamnose, ribose, glucose, galactose, and mannose. The wall material that was released contained only 47% amino acids and was enriched in lipid, glucose, and phosphate. These results support data from electron micrographs, showing the localized release of cell wall material by the disulfide bond-breaking reagent at alkaline pH. In amino acid composition the untreated walls did not differ greatly from the material released by dithiothreitol, but differed considerably from the walls of another strain of M. hungatii. The ratios of the amino acids found in the cell wall proteins of several archaebacteria and of Bacillus cereus spore coats were similar.

scholarly journals Cytokinesis in fra2 Arabidopsis thaliana p60-Katanin Mutant: Defects in Cell Plate/Daughter Wall Formation

2021 ◽  
Vol 22 (3) ◽  
pp. 1405
Author(s):  
Emmanuel Panteris ◽  
Anna Kouskouveli ◽  
Dimitris Pappas ◽  
Ioannis-Dimosthenis S. Adamakis
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Cell Walls ◽  
Higher Plants ◽  
Cell Plate ◽  
Wild Type ◽  
Microtubule Organization ◽  
Root Cells ◽  
Wall Formation ◽  
In The Wild

Cytokinesis is accomplished in higher plants by the phragmoplast, creating and conducting the cell plate to separate daughter nuclei by a new cell wall. The microtubule-severing enzyme p60-katanin plays an important role in the centrifugal expansion and timely disappearance of phragmoplast microtubules. Consequently, aberrant structure and delayed expansion rate of the phragmoplast have been reported to occur in p60-katanin mutants. Here, the consequences of p60-katanin malfunction in cell plate/daughter wall formation were investigated by transmission electron microscopy (TEM), in root cells of the fra2 Arabidopsis thaliana loss-of-function mutant. In addition, deviations in the chemical composition of cell plate/new cell wall were identified by immunolabeling and confocal microscopy. It was found that, apart from defective phragmoplast microtubule organization, cell plates/new cell walls also appeared faulty in structure, being unevenly thick and perforated by large gaps. In addition, demethylesterified homogalacturonans were prematurely present in fra2 cell plates, while callose content was significantly lower than in the wild type. Furthermore, KNOLLE syntaxin disappeared from newly formed cell walls in fra2 earlier than in the wild type. Taken together, these observations indicate that delayed cytokinesis, due to faulty phragmoplast organization and expansion, results in a loss of synchronization between cell plate growth and its chemical maturation.

scholarly journals Plant cell wall integrity maintenance and pattern-triggered immunity modulate jointly plant stress responses in Arabidopsis thaliana

2017 ◽  
Author(s):  
Timo Engelsdorf ◽  
Nora Gigli-Bisceglia ◽  
Manikandan Veerabagu ◽  
Joseph F. McKenna ◽  
Frauke Augstein ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Plant Growth ◽  
Plant Cell ◽  
Stress Responses ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Cell Wall Integrity ◽  
Maintenance Mechanism ◽  
Signaling Components

AbstractPlant cells are surrounded by walls, which must often meet opposing functional requirements during plant growth and defense. The cells meet them by modifying wall structure and composition in a tightly controlled and adaptive manner. The modifications seem to be mediated by a dedicated cell wall integrity (CWI) maintenance mechanism. Currently the mode of action of the mechanism is not understood and it is unclear how its activity is coordinated with established plant defense signaling. We investigated responses to induced cell wall damage (CWD) impairing CWI and the underlying mechanism in Arabidopsis thaliana. Interestingly inhibitor- and enzyme-derived CWD induced similar, turgor-sensitive stress responses. Genetic analysis showed that the receptor-like kinase (RLK) FEI2 and the mechano-sensitive, plasma membrane-localized Ca2+- channel MCA1 function downstream of the THE1 RLK in CWD perception. Phenotypic clustering with 27 genotypes identified a core group of RLKs and ion channels, required for activation of CWD responses. By contrast, the responses were repressed by pattern-triggered immune (PTI) signaling components including PEPR1 and 2, the receptors for the immune signaling peptide AtPep1. Interestingly AtPep1 application repressed CWD-induced phytohormone accumulation in a PEPR1/2-dependent manner. These results suggest that PTI suppresses CWD-induced defense responses through elicitor peptide-mediated signaling during defense response activation. If PTI is impaired, the suppression of CWD-induced responses is alleviated, thus compensating for defective PTI.Significance statementStress resistance and plant growth determine food crop yield and efficiency of bioenergy production from ligno-cellulosic biomass. Plant cell walls are essential elements of the biological processes, therefore functional integrity of the cell walls must be maintained throughout. Here we investigate the plant cell wall integrity maintenance mechanism. We characterize its mode of action, identify essential signaling components and show that the AtPep1 signaling peptide apparently coordinates pattern triggered immunity (PTI) and cell wall integrity maintenance in plants. These results suggest how PTI and cell wall modification coordinately regulate biotic stress responses with plants possibly compensating for PTI impairment through enhanced activation of stress responses regulated by the CWI maintenance mechanism.

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