Immunogold localization of callose and other plant cell wall components in soybean roots infected with the oomycete Phytophthora sojae

1997 ◽  
Vol 75 (9) ◽  
pp. 1509-1517 ◽  
Author(s):  
K. Enkerli ◽  
C. W. Mims ◽  
M. G. Hahn
Keyword(s):  
Plasma Membrane ◽  
Plant Cell Wall ◽  
Arabinogalactan Proteins ◽  
Phytophthora Sojae ◽  
Arabinogalactan Protein ◽  
Electron Microscopic ◽  
Rhamnogalacturonan I ◽  
Host Plasma Membrane ◽  
Wall Appositions ◽  
Extrahaustorial Matrix

Immunolabeling and transmission electron microscopic techniques were used to investigate the chemical nature of wall appositions in roots of susceptible and resistant soybean plants inoculated with Phytophthora sojae race 2. The extrahaustorial matrix associated with the haustorium of Phytophthora sojae also was examined. Antibodies against (1 → 3)-β-glucan, a terminal α-fucosyl-containing epitope present in xyloglucan and rhamnogalacturonan I, and an arabinosylated (1 → 6)-β-galactan epitope present in arabinogalactan proteins were used. (1 → 3)-β-Glucan (callose), xyloglucan, and arabinogalactan proteins were found to be localized in all wall appositions regardless of how long after inoculation the appositions developed or whether plants were susceptible or resistant to Phytophthora sojae. (1 → 3)-β-Glucan also was found in fungal walls and at host cell plasmodesmata. None of the four antibodies labeled the extrahaustorial matrix. The antibody against arabinogalactan protein recognized the host plasma membrane, but not the invaginated host plasma membrane associated with the extrahaustorial matrix. This result indicates that the properties or the composition of the host plasma membrane may change locally once it becomes an extrahaustorial membrane. Key words: Phytophthora sojae, Glycine max, callose, immunolabeling, wall appositions, papillae.

scholarly journals Arabinogalactan protein–rare earth element complexes activate plant endocytosis

2019 ◽  
Vol 116 (28) ◽  
pp. 14349-14357 ◽  
Author(s):  
Lihong Wang ◽  
Mengzhu Cheng ◽  
Qing Yang ◽  
Jigang Li ◽  
Xiang Wang ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Earth Element ◽  
Adaptor Protein ◽  
Arabinogalactan Proteins ◽  
Arabinogalactan Protein ◽  
Outer Face ◽  
Transmembrane Receptors ◽  
Superresolution Imaging ◽  
Endocytic Vesicles ◽  
Current Paradigm

Endocytosis is essential to all eukaryotes, but how cargoes are selected for internalization remains poorly characterized. Extracellular cargoes are thought to be selected by transmembrane receptors that bind intracellular adaptors proteins to initiate endocytosis. Here, we report a mechanism for clathrin-mediated endocytosis (CME) of extracellular lanthanum [La(III)] cargoes, which requires extracellular arabinogalactan proteins (AGPs) that are anchored on the outer face of the plasma membrane. AGPs were colocalized with La(III) on the cell surface and in La(III)-induced endocytic vesicles inArabidopsisleaf cells. Superresolution imaging showed that La(III) triggered AGP movement across the plasma membrane. AGPs were then colocalized and physically associated with the μ subunit of the intracellular adaptor protein 2 (AP2) complexes. The AGP-AP2 interaction was independent of CME, whereas AGP’s internalization required CME and AP2. Moreover, we show that AGP-dependent endocytosis in the presence of La(III) also occurred in human cells. These findings indicate that extracellular AGPs act as conserved CME cargo receptors, thus challenging the current paradigm about endocytosis of extracellular cargoes.

An ultrastructural study of the marine diatom Licmophora hyalina and its parasite Ectrogella perforons. I. Infection of host cells

1980 ◽  
Vol 58 (11) ◽  
pp. 1280-1290 ◽  
Author(s):  
Chandralata Raghu Kumar
Keyword(s):  
Plasma Membrane ◽  
Germ Tube ◽  
Host Cells ◽  
Marine Diatom ◽  
Electron Microscopic ◽  
Fungal Parasite ◽  
Successful Infection ◽  
Host Cell Wall ◽  
Host Plasma Membrane ◽  
Host Parasite

An electron microscopic study has been made on the infection and penetration of the marine diatom Licmophora hyalina Agardh by Ectrogella perforons Petersen, an obligate fungal parasite of diatoms. The zoospores encyst on the host cell wall. The nucleus of the cyst may be situated proximal or distal to the host wall. A germ tube is produced from the side where the nucleus is situated. The germ tube may be branched or unbranched. The penetrating germ tube swells distally, develops an appressorium at the site of penetration of the host wall, and pierces the host wall in the form of an infection peg. The infection peg is smaller in diameter than the germ tube and the appressorium. Successful infection takes place always at the areolae of the diatom wall. The infection peg may directly inject its contents by piercing the subfrustular layer of the diatom wall or may grow for some distance beneath the subfrustular layer. At the site of entry the host plasma membrane invaginates and surrounds the fungal protoplast. Initially, the host–parasite interface consists of a two-layered envelope of which the outer one is the host plasma membrane and the inner one the fungal plasma membrane.

scholarly journals Structural Basis of the Function of Yariv Reagent—An Important Tool to Study Arabinogalactan Proteins

2021 ◽  
Vol 8 ◽  
Author(s):  
Tereza Přerovská ◽  
Anna Pavlů ◽  
Dzianis Hancharyk ◽  
Anna Rodionova ◽  
Anna Vavříková ◽  
...  
Keyword(s):  
Plant Cell Wall ◽  
Plant Defence ◽  
Three Dimensional ◽  
Arabinogalactan Proteins ◽  
Arabinogalactan Protein ◽  
Dimensional Structure ◽  
Structural Basis ◽  
Functional Studies ◽  
Yariv Reagent ◽  
Dynamics Simulations

Arabinogalactan proteins are very abundant, heavily glycosylated plant cell wall proteins. They are intensively studied because of their crucial role in plant development as well as their function in plant defence. Research of these biomacromolecules is complicated by the lack of tools for their analysis and characterisation due to their extreme heterogeneity. One of the few available tools for detection, isolation, characterisation, and functional studies of arabinogalactan proteins is Yariv reagents. Yariv reagent is a synthetic aromatic glycoconjugate originally prepared as an antigen for immunization. Later, it was found that this compound can precipitate arabinogalactan proteins, namely, their ß-D-(1→3)-galactan structures. Even though this compound has been intensively used for decades, the structural basis of arabinogalactan protein precipitation by Yariv is not known. Multiple biophysical studies have been published, but none of them attempted to elucidate the three-dimensional structure of the Yariv-galactan complex. Here we use a series of molecular dynamics simulations of systems containing one or multiple molecules of ß-D-galactosyl Yariv reagent with or without oligo ß-D-(1→3)-galactan to predict the structure of the complex. According to our model of Yariv-galactan complexes, Yariv reagent forms stacked oligomers stabilized by π-π and CH/π interactions. These oligomers may contain irregularities. Galactan structures crosslink these Yariv oligomers. The results were compared with studies in literature.

Immunocytochemical localization of p65 with one-nanometer gold particles following silver development

1989 ◽  
Vol 47 ◽  
pp. 1042-1043
Author(s):  
Richard W. Burry ◽  
Diane M. Hayes
Keyword(s):  
Plasma Membrane ◽  
Bovine Serum Albumin ◽  
Calf Serum ◽  
Specific Protein ◽  
Immunocytochemical Localization ◽  
Electron Microscopic ◽  
Gold Particles ◽  
Pass Through ◽  
Label Distribution ◽  
Phosphate Buffered Saline

Electron microscopic (EM) immunocytochemistry localization of the neuron specific protein p65 could show which organelles contain this antigen. Antibodies (Ab) labeled with horseradish peroxidase (HRP) followed by chromogen development show a broad diffuse label distribution within cells and restricting identification of organelles. Particulate label (e.g. 10 nm colloidal gold) is highly desirable but not practical because penetration into cells requires destroying the plasma membrane. We report pre-embedding immunocytochemistry with a particulate marker, 1 nm gold, that will pass through membranes treated with saponin, a mild detergent.Cell cultures of the rat cerebellum were fixed in buffered 4% paraformaldehyde and 0.1% glutaraldehyde (Glut.). The buffer for all incubations and rinses was phosphate buffered saline with: 1% calf serum, 0.2% saponin, 0.1% gelatin, 50 mM glycine 1 mg/ml bovine serum albumin, and (not in the HRP labeled cultures) 0.02% sodium azide. The monoclonal #48 to p65 was used with three label systems: HRP, 1 nm avidin gold with IntenSE M development, and 1 nm avidin gold with Danscher development.

scholarly journals Electron Microscopic Study of the Phagocytosis Process in Lung

1960 ◽  
Vol 7 (2) ◽  
pp. 357-366 ◽  
Author(s):  
H. E. Karrer
Keyword(s):  
Plasma Membrane ◽  
Alveolar Macrophages ◽  
Alveolar Epithelium ◽  
Alveolar Epithelial Cells ◽  
Electron Microscopic ◽  
Double Line ◽  
Alveolar Epithelial ◽  
Culture Cells ◽  
India Ink ◽  
Intracellular Vesicles

Diluted India ink was instilled into the nasal cavity of mice and the lungs of some animals were fixed with osmium tetroxide at various intervals after one instillation. The lungs of other animals were fixed after 4, 7, 9, 16, or 18 daily instillations. The India ink was found to be phagocytized almost exclusively by the free alveolar macrophages. A few particles are occasionally seen within thin portions of alveolar epithelium, within the "small" alveolar epithelial cells, or within occasional leukocytes in the lumina of alveoli. The particles are ingested by an invagination process of the plasma membrane resulting in the formation of intracellular vesicles and vacuoles. Ultimately large amounts of India ink accumulate in the cell, occupying substantial portions of the cytoplasm. The surfaces of phagocytizing macrophages show signs of intense motility. Their cytoplasm contains numerous particles, resembling Palade particles, and a large amount of rough surfaced endoplasmic reticulum. These structures are interpreted as indicative of protein synthesis. At the level of resolution achieved in this study the membranes of this reticulum appear as single dense "lines." On the other hand, the plasma membrane and the limiting membranes of vesicles and of vacuoles often exhibit the double-line structure typical of unit membranes (Robertson, 37). The inclusion bodies appear to be the product of phagocytosis. It is believed that some of them derive from the vacuoles mentioned above, and that they correspond to similar structures seen in phase contrast cinemicrographs of culture cells. Their matrix represents phagocytized material. Certain structures within this matrix are considered as secondary and some of these structures possess an ordered form probably indicative of the presence of lipid. The possible origin and the fate of alveolar macrophages are briefly discussed.

scholarly journals PROBLEMS ASSOCIATED WITH THE DEMONSTRATION BY LEAD METHODS OF ADENOSINE TRIPHOSPHATASE ACTIVITY IN RESIDENT PERITONEAL MACROPHAGES AND EXUDATE MONOCYTES OF THE GUINEA PIG

1973 ◽  
Vol 21 (5) ◽  
pp. 488-498 ◽  
Author(s):  
R. E. POELMANN ◽  
W. T. DAEMS ◽  
E. J. VAN LOHUIZEN
Keyword(s):  
Plasma Membrane ◽  
Guinea Pig ◽  
Microscopic Study ◽  
Heterogeneous Nucleation ◽  
Adenosine Triphosphatase ◽  
Peritoneal Macrophages ◽  
Electron Microscopic ◽  
Adenosine Triphosphatase Activity ◽  
Eosinophilic Granulocytes ◽  
Membrane Bound

This cytochemical and electron microscopic study on peritoneal macrophages of the guinea pig has raised doubts concerning the validity of lead methods for the demonstration of plasma membrane-bound adenosine triphosphatase activity. The problems encountered are inherent in the use of lead ions as a capture reagent. The nonenzymatically formed precipitates reflect sites of heterogeneous nucleation specific for certain kinds of cells, e.g., resident peritoneal macrophages, eosinophilic granulocytes and, to a lesser degree, exudate monocytes. This type of precipitation is also catalyzed on the surface of nonbiologic matrices such as latex particles. Enzymatic processes may well occur, but they cannot be distinguished from nonenzymatic processes.

Etude ultrastructurale des relations hôte–parasite au cours de l'infection des pommes par le Phytophthora cactorum

1981 ◽  
Vol 59 (2) ◽  
pp. 251-263 ◽  
Author(s):  
X. Mourichon ◽  
G. Sallé
Keyword(s):  
Cell Walls ◽  
Susceptible Variety ◽  
Host Cells ◽  
Phytophthora Cactorum ◽  
Electron Microscopic ◽  
Susceptible Host ◽  
Golden Delicious ◽  
Extrahaustorial Matrix ◽  
Apple Fruits

An electron microscopic study was performed on haustoria of Phytophthora cactorum (L. et C.) Schroeter developed in tissues of two cultivars of apple fruits: a susceptible variety ('Golden delicious') and a resistant one ('Belle de Boskoop'). Ultrastructure of intercellular hyphae and some aspects of their penetration between contiguous host cells were described. A light dissolution of the host cell walls was observed. Ontogenic investigations indicated that in the susceptible host, the wall of the fungal haustoria was covered with a dense-stained extrahaustorial matrix. Its origin and its polysaccharide nature were demonstrated. On the other hand, the resistant host developed, immediately after the inoculation, a papilla which gave rise, later on, to a sheath enclosing adult haustoria. The role of these callosic structures in the phenomenon of resistance was discussed.

Lipocortin 1 (annexin 1) in patches associated with the membrane of a lung adenocarcinoma cell line and in the cell cytoplasm

1998 ◽  
Vol 111 (10) ◽  
pp. 1405-1418 ◽  
Author(s):  
V. Traverso ◽  
J.F. Morris ◽  
R.J. Flower ◽  
J. Buckingham
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Lung Adenocarcinoma ◽  
Western Blotting ◽  
Signal Sequence ◽  
Membrane Surface ◽  
Integral Membrane Protein ◽  
Subcellular Fractionation ◽  
Cell Fractionation ◽  
Electron Microscopic

Lipocortin 1 (annexin I) is a calcium- and phospholipid-binding annexin protein which can be externalised from cells despite the lack of a signal sequence. To determine its cellular distribution lipocortin 1 in A549 human lung adenocarcinoma cells was localised by light- and electron-microscopic immunocytochemistry and by cell fractionation and western blotting. Lipocortin 1 immunoreactivity is concentrated in prominent patches associated with the plasma membrane. The intensity of these patches varied with the confluence and duration of the culture and was not detectably diminished by an EDTA wash before fixation. Tubulin and cytokeratin 8 were colocalized with lipocortin 1 in the patches. Within the cells lipocortin 1 was distributed throughout the cytoplasm. Electron microscopy revealed prominent immunoreactivity along the plasma membrane with occasional large clusters of gold particles in contact with the membrane surface of the cells; within the cytoplasm the membrane of some vesicle/vacuole structures and some small electron-dense bodies was immunoreactive, but no immunogold particles were associated with the multilamellar bodies. Subcellular fractionation, extraction and western blotting showed that lipocortin 1 in the membrane pellet was present as two distinct fractions; one, intimately associated with the lipid bilayer, which behaved like an integral membrane protein and one loosely attached which behaved like a peripheral membrane protein. The results show that a substantial amounts of lipocortin 1 is concentrated in focal structures associated with and immediately beneath the plasma membrane. These might form part of the mechanism by which lipocortin 1 is released from the cells.

scholarly journals GRAMP 100: a membrane protein concentrated on secretory granule membranes and the apical cell surface in exocrine acinar cells.

1992 ◽  
Vol 40 (12) ◽  
pp. 1827-1835 ◽  
Author(s):  
S M Laurie ◽  
M B Mixon ◽  
J D Castle
Keyword(s):  
Plasma Membrane ◽  
Apical Cell ◽  
Reduced Form ◽  
Electron Microscopic Level ◽  
Apical Plasma Membrane ◽  
Cell Fractionation ◽  
Immunocytochemical Localization ◽  
Electron Microscopic ◽  
Common Component ◽  
Rat Parotid

Using a monoclonal antibody (SG10A6) raised against secretion granule membranes of the rat parotid gland, we have identified an antigen that is a common component of both exocrine pancreatic and parotid granule membranes. SG10A6 (an IgM) immunoprecipitates antigen that migrates as a single band (M(r) approximately 80 KD unreduced; M(r) approximately 100 KD reduced) and immunoblots at least two polypeptides that are similar to the reduced and nonreduced immunoprecipitated antigen. This granule-associated membrane polypeptide (GRAMP 100; named for the apparent M(r) in reduced form) is also a prominent component of plasma membrane fractions. Immunocytochemical localization at the electron microscopic level demonstrates the presence of GRAMP 100 on granule membranes, especially condensing vacuoles and exocytotic figures, and the apical plasma membrane. Lower levels of antigen are detected on basolateral plasma membrane and on peri-Golgi membranes that may be part of the endosomal system. Both the cell fractionation and immunocytochemical localization indicate that GRAMP 100 differs in distribution from GRAMP 92 and 30K SCAMPs, two other components of exocrine granule membranes identified with monoclonal antibodies. To date, no polypeptides have been identified with this approach that are exclusive components of exocrine granule membranes.

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