scholarly journals Structural studies on cell wall biogenesis-I: A method to produce high-quality protoplasts from alga Micrasterias denticulata, an emerging model in plant biology

2021 ◽  
Author(s):  
M Selvaraj
Keyword(s):  
Cell Wall ◽  
Cell Biology ◽  
Model System ◽  
Plant Biology ◽  
Cell Wall Biogenesis ◽  
Unicellular Green Alga ◽  
Biological Studies ◽  
Micrasterias Denticulata ◽  
And Behavior ◽  
Protoplast Production

Micrasterias denticulate is a freshwater unicellular green alga emerging as a model system in plant cell biology. This is an algae that has been examined in the context of cell wall research from early 1970s. Protoplast production from such a model system is important for many downstream physiological and cell biological studies. The algae produce intact protoplast in a straight two-step protocol involving 5% mannitol, 2% cellulysin, 4mM calcium chloride under a temperature ramping strategy. The process of protoplast induction and behavior of protoplast was examined by light microscopy and reported in this study.

scholarly journals The effects of applied electric fields on Micrasterias. I. Morphogenesis and the pattern of cell wall deposition

1980 ◽  
Vol 42 (1) ◽  
pp. 261-277
Author(s):  
D.L. Brower ◽  
J.R. McIntosh
Keyword(s):  
Cell Wall ◽  
Electric Fields ◽  
Cell Expansion ◽  
Applied Field ◽  
Potential Drop ◽  
Wall Material ◽  
Methyl Groups ◽  
Unicellular Green Alga ◽  
Micrasterias Denticulata ◽  
Cell Wall Deposition

Applied electric fields of approximately 14 V/cm have profound effects on the morphogenesis of the unicellular green alga, Micrasterias denticulata. This field corresponds to a potential drop of 15–40 mV across lobes oriented perpendicular to the applied field. These lobes show a galvanotropism toward the cathode. Lobes growing parallel to the field are stunted to varying degrees, depending on their orientation. As shown by other investigators, most cell wall material is normally deposited at the tips of growing lobes. If, however, cell expansion is osmotically inhibited in electric fields, wall material also accumulates along the cathode-facing (CF) sides of lobes oriented perpendicular to the field. Similarly, in cells growing under the influence of an applied field, radioactively labelled glucose and the methyl groups from methionine are incorporated along the CF sides of lobes as well as the lobe tips. This is also true when the label is added immediately after cells are removed from the fields, indicating that the wall-depositing machinery itself has been temporarily altered by the field. These results demonstrate that applied electric fields can be a valuable tool in elucidating the mechanisms of growth localization in Micrasterias cells.

scholarly journals Pathway-Directed Screen for Inhibitors of the Bacterial Cell Elongation Machinery

2018 ◽  
Vol 63 (1) ◽  
Author(s):  
Jackson A. Buss ◽  
Vadim Baidin ◽  
Michael A. Welsh ◽  
Josué Flores-Kim ◽  
Hongbaek Cho ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Elongation ◽  
Bacterial Infections ◽  
Strongly Correlated ◽  
Beta Lactam ◽  
Content Type ◽  
Cell Wall Biogenesis ◽  
Biological Studies ◽  
Rod System ◽  
New Antibiotics

ABSTRACT New antibiotics are needed to combat the growing problem of resistant bacterial infections. An attractive avenue toward the discovery of such next-generation therapies is to identify novel inhibitors of clinically validated targets, like cell wall biogenesis. We have therefore developed a pathway-directed whole-cell screen for small molecules that block the activity of the Rod system of Escherichia coli. This conserved multiprotein complex is required for cell elongation and the morphogenesis of rod-shaped bacteria. It is composed of cell wall synthases and membrane proteins of unknown function that are organized by filaments of the actin-like MreB protein. Our screen takes advantage of the conditional essentiality of the Rod system and the ability of the beta-lactam mecillinam (also known as amdinocillin) to cause a toxic malfunctioning of the machinery. Rod system inhibitors can therefore be identified as molecules that promote growth in the presence of mecillinam under conditions permissive for the growth of Rod– cells. A screen of ∼690,000 compounds identified 1,300 compounds that were active against E. coli. Pathway-directed screening of a majority of this subset of compounds for Rod inhibitors successfully identified eight analogs of the MreB antagonist A22. Further characterization of the A22 analogs identified showed that their antibiotic activity under conditions where the Rod system is essential was strongly correlated with their ability to suppress mecillinam toxicity. This result combined with those from additional biological studies reinforce the notion that A22-like molecules are relatively specific for MreB and suggest that the lipoprotein transport factor LolA is unlikely to be a physiologically relevant target as previously proposed.

New insights on Laminaria digitata ultrastructure through combined conventional chemical fixation and cryofixation

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Christos Katsaros ◽  
Sophie Le Panse ◽  
Gillian Milne ◽  
Carl J. Carrano ◽  
Frithjof Christian Küpper
Keyword(s):  
Cell Wall ◽  
General Structure ◽  
Raw Material ◽  
Rocky Shores ◽  
Chemical Fixation ◽  
Laminaria Digitata ◽  
Vegetative Cells ◽  
Biological Studies ◽  
New Findings ◽  
Wide Range

Abstract The objective of the present study is to examine the fine structure of vegetative cells of Laminaria digitata using both chemical fixation and cryofixation. Laminaria digitata was chosen due to its importance as a model organism in a wide range of biological studies, as a keystone species on rocky shores of the North Atlantic, its use of iodide as a unique inorganic antioxidant, and its significance as a raw material for the production of alginate. Details of the fine structural features of vegetative cells are described, with particular emphasis on the differences between the two methods used, i.e. conventional chemical fixation and freeze-fixation. The general structure of the cells was similar to that already described, with minor differences between the different cell types. An intense activity of the Golgi system was found associated with the thick external cell wall, with large dictyosomes from which numerous vesicles and cisternae are released. An interesting type of cisternae was found in the cryofixed material, which was not visible with the chemical fixation. These are elongated structures, in sections appearing tubule-like, close to the external cell wall or to young internal walls. An increased number of these structures was observed near the plasmodesmata of the pit fields. They are similar to the “flat cisternae” found associated with the forming cytokinetic diaphragm of brown algae. Their possible role is discussed. The new findings of this work underline the importance of such combined studies which reveal new data not known until now using the old conventional methods. The main conclusion of the present study is that cryofixation is the method of choice for studying Laminaria cytology by transmission electron microscopy.

scholarly journals Microtubule Dynamics Plays a Vital Role in Plant Adaptation and Tolerance to Salt Stress

2021 ◽  
Vol 22 (11) ◽  
pp. 5957
Author(s):  
Hyun Jin Chun ◽  
Dongwon Baek ◽  
Byung Jun Jin ◽  
Hyun Min Cho ◽  
Mi Suk Park ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Wall ◽  
Salt Stress ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Related Gene ◽  
Plant Adaptation ◽  
Salt Stress Response ◽  
Network Analyses ◽  
Cell Wall Biogenesis

Although recent studies suggest that the plant cytoskeleton is associated with plant stress responses, such as salt, cold, and drought, the molecular mechanism underlying microtubule function in plant salt stress response remains unclear. We performed a comparative proteomic analysis between control suspension-cultured cells (A0) and salt-adapted cells (A120) established from Arabidopsis root callus to investigate plant adaptation mechanisms to long-term salt stress. We identified 50 differentially expressed proteins (45 up- and 5 down-regulated proteins) in A120 cells compared with A0 cells. Gene ontology enrichment and protein network analyses indicated that differentially expressed proteins in A120 cells were strongly associated with cell structure-associated clusters, including cytoskeleton and cell wall biogenesis. Gene expression analysis revealed that expressions of cytoskeleton-related genes, such as FBA8, TUB3, TUB4, TUB7, TUB9, and ACT7, and a cell wall biogenesis-related gene, CCoAOMT1, were induced in salt-adapted A120 cells. Moreover, the loss-of-function mutant of Arabidopsis TUB9 gene, tub9, showed a hypersensitive phenotype to salt stress. Consistent overexpression of Arabidopsis TUB9 gene in rice transgenic plants enhanced tolerance to salt stress. Our results suggest that microtubules play crucial roles in plant adaptation and tolerance to salt stress. The modulation of microtubule-related gene expression can be an effective strategy for developing salt-tolerant crops.

scholarly journals Fundamental functional differences between gyri and sulci: implications for brain function, cognition, and behavior

2021 ◽  
Vol 1 (1) ◽  
pp. 23-41
Author(s):  
Xi Jiang ◽  
Tuo Zhang ◽  
Shu Zhang ◽  
Keith M Kendrick ◽  
Tianming Liu
Keyword(s):  
Brain Function ◽  
Cell Biology ◽  
Functional Anatomy ◽  
Building Blocks ◽  
Functional Differentiation ◽  
Cortical Folding ◽  
Brain Functions ◽  
Functional Relationships ◽  
Structural Architecture ◽  
And Behavior

Abstract Folding of the cerebral cortex is a prominent characteristic of mammalian brains. Alterations or deficits in cortical folding are strongly correlated with abnormal brain function, cognition, and behavior. Therefore, a precise mapping between the anatomy and function of the brain is critical to our understanding of the mechanisms of brain structural architecture in both health and diseases. Gyri and sulci, the standard nomenclature for cortical anatomy, serve as building blocks to make up complex folding patterns, providing a window to decipher cortical anatomy and its relation with brain functions. Huge efforts have been devoted to this research topic from a variety of disciplines including genetics, cell biology, anatomy, neuroimaging, and neurology, as well as involving computational approaches based on machine learning and artificial intelligence algorithms. However, despite increasing progress, our understanding of the functional anatomy of gyro-sulcal patterns is still in its infancy. In this review, we present the current state of this field and provide our perspectives of the methodologies and conclusions concerning functional differentiation between gyri and sulci, as well as the supporting information from genetic, cell biology, and brain structure research. In particular, we will further present a proposed framework for attempting to interpret the dynamic mechanisms of the functional interplay between gyri and sulci. Hopefully, this review will provide a comprehensive summary of anatomo-functional relationships in the cortical gyro-sulcal system together with a consideration of how these contribute to brain function, cognition, and behavior, as well as to mental disorders.

scholarly journals The Transcription Factor Con7-1 Is a Master Regulator of Morphogenesis and Virulence in Fusarium oxysporum

2015 ◽  
Vol 28 (1) ◽  
pp. 55-68 ◽  
Author(s):  
Carmen Ruiz-Roldán ◽  
Yolanda Pareja-Jaime ◽  
José Antonio González-Reyes ◽  
M. Isabel G. Roncero
Keyword(s):  
Cell Wall ◽  
Fusarium Oxysporum ◽  
Gene Expression Analysis ◽  
Wall Structure ◽  
Targeted Deletion ◽  
Signal Perception ◽  
Cell Wall Biogenesis ◽  
Primary And Secondary Metabolism ◽  
Genes Encoding

Previous studies have demonstrated the essential role of morphogenetic regulation in Fusarium oxysporum pathogenesis, including processes such as cell-wall biogenesis, cell division, and differentiation of infection-like structures. We identified three F. oxysporum genes encoding predicted transcription factors showing significant identities to Magnaporthe oryzae Con7p, Con7-1, plus two identical copies of Con7-2. Targeted deletion of con7-1 produced nonpathogenic mutants with altered morphogenesis, including defects in cell wall structure, polar growth, hyphal branching, and conidiation. By contrast, simultaneous inactivation of both con7-2 copies caused no detectable defects in the resulting mutants. Comparative microarray-based gene expression analysis indicated that Con7-1 modulates the expression of a large number of genes involved in different biological functions, including host–pathogen interactions, morphogenesis and development, signal perception and transduction, transcriptional regulation, and primary and secondary metabolism. Taken together, our results point to Con7-1 as general regulator of morphogenesis and virulence in F. oxysporum.

scholarly journals Reconstruction ofmreBExpression in Staphylococcus aureus via a Collection of New Integrative Plasmids

2014 ◽  
Vol 80 (13) ◽  
pp. 3868-3878 ◽  
Author(s):  
Ana Yepes ◽  
Gudrun Koch ◽  
Andrea Waldvogel ◽  
Juan-Carlos Garcia-Betancur ◽  
Daniel Lopez
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Cell Biology ◽  
Genetic Manipulation ◽  
Protein Localization ◽  
Antibiotic Resistance Genes ◽  
Wall Material ◽  
Content Type ◽  
Genetic Manipulations ◽  
Discrete Structures

ABSTRACTProtein localization has been traditionally explored in unicellular organisms, whose ease of genetic manipulation facilitates molecular characterization. The two rod-shaped bacterial modelsEscherichia coliandBacillus subtilishave been prominently used for this purpose and have displaced other bacteria whose challenges for genetic manipulation have complicated any study of cell biology. Among these bacteria is the spherical pathogenic bacteriumStaphylococcus aureus. In this report, we present a new molecular toolbox that facilitates gene deletion in staphylococci in a 1-step recombination process and additional vectors that facilitate the insertion of diverse reporter fusions into newly identified neutral loci of theS. aureuschromosome. Insertion of the reporters does not add any antibiotic resistance genes to the chromosomes of the resultant strains, thereby making them amenable for further genetic manipulations. We used this toolbox to reconstitute the expression ofmreBinS. aureus, a gene that encodes an actin-like cytoskeletal protein which is absent in coccal cells and is presumably lost during the course of speciation. We observed that inS. aureus, MreB is organized in discrete structures in association with the membrane, leading to an unusual redistribution of the cell wall material. The production of MreB also caused cell enlargement, but it did not revert staphylococcal shape. We present interactions of MreB with key staphylococcal cell wall-related proteins. This work facilitates the useS. aureusas a model system in exploring diverse aspects of cellular microbiology.

scholarly journals The Cek1 and Hog1 Mitogen-Activated Protein Kinases Play Complementary Roles in Cell Wall Biogenesis and Chlamydospore Formation in the Fungal Pathogen Candida albicans

2006 ◽  
Vol 5 (2) ◽  
pp. 347-358 ◽  
Author(s):  
B. Eisman ◽  
R. Alonso-Monge ◽  
E. Román ◽  
D. Arana ◽  
C. Nombela ◽  
...  
Keyword(s):  
Cell Wall ◽  
Candida Albicans ◽  
Map Kinase ◽  
Fungal Pathogen ◽  
Hog Pathway ◽  
Cell Wall Biogenesis ◽  
Chlamydospore Formation ◽  
Mitogen Activated Protein ◽  
Morphological Transitions ◽  
Relationship Of

ABSTRACT The Hog1 mitogen-activated protein (MAP) kinase mediates an adaptive response to both osmotic and oxidative stress in the fungal pathogen Candida albicans. This protein also participates in two distinct morphogenetic processes, namely the yeast-to-hypha transition (as a repressor) and chlamydospore formation (as an inducer). We show here that repression of filamentous growth occurs both under serum limitation and under other partially inducing conditions, such as low temperature, low pH, or nitrogen starvation. To understand the relationship of the HOG pathway to other MAP kinase cascades that also play a role in morphological transitions, we have constructed and characterized a set of double mutants in which we deleted both the HOG1 gene and other signaling elements (the CST20, CLA4, and HST7 kinases, the CPH1 and EFG1 transcription factors, and the CPP1 protein phosphatase). We also show that Hog1 prevents the yeast-to-hypha switch independent of all the elements analyzed and that the inability of the hog1 mutants to form chlamydospores is suppressed when additional elements of the CEK1 pathway (CST20 or HST7) are altered. Finally, we report that Hog1 represses the activation of the Cek1 MAP kinase under basal conditions and that Cek1 activation correlates with resistance to certain cell wall inhibitors (such as Congo red), demonstrating a role for this pathway in cell wall biogenesis.

Sign in / Sign up

Export Citation Format

Share Document