Light-induced conidiation ofTrichodermaspp. strains is accompanied by development-dependent changes in the Ca2+binding to cell walls

2018 ◽  
Vol 64 (11) ◽  
pp. 856-864 ◽  
Author(s):  
Szilvia Kontár ◽  
L’udovít Varečka ◽  
Michal Híreš ◽  
Svetlana Kryštofová ◽  
Martin Šimkovič
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Walls ◽  
Binding Capacity ◽  
Trichoderma Atroviride ◽  
Major Fraction ◽  
Trichoderma Spp ◽  
Intracellular Compartments ◽  
Effect Of Light

The effect of light on the binding of Ca2+to mycelia and to cell walls isolated from aerial mycelia of three strains of Trichoderma spp. was studied. Two independent methods were used to measure the total Ca2+content in mycelia and the Ca2+bound to cell walls isolated from aerial mycelia. The results of these methods showed that the light-induced formation and maturation of conidia in Trichoderma spp. is accompanied by increased Ca2+deposition in mycelia and cell walls. Moreover, the cultivation of Trichoderma atroviride F-534 in the presence of45Ca2+under circadian illumination showed that radioactivity was exclusively localized in the light-induced conidial rings of aerial mycelia. The fluorescence microscopy of chlortetracycline-stained mycelia showed that the major fraction of Ca2+was accumulated in conidia and fructification structures, or some intracellular compartments in T. atroviride F-534 grown under circadian illumination, while only a limited amount of Ca2+was associated with hyphal surfaces. In addition, the study of45Ca2+binding to cell walls revealed that T. atroviride F-534 displays both increased45Ca2+binding capacity and elevated affinity to45Ca2+binding upon illumination. The results indicate that conidia formation and (or) maturation is associated with changes in Ca2+homeostasis.

Internalisation of fluorescein isothiocyanate and fluorescein isothiocyanatedextran by suspension-cultured plant cells

1990 ◽  
Vol 96 (4) ◽  
pp. 721-730
Author(s):  
LOUISE COLE ◽  
JULLIAN COLEMAN ◽  
DAVID EVANS ◽  
CHRIS HAWES
Keyword(s):  
Fluorescence Microscopy ◽  
Incubation Period ◽  
Degradation Products ◽  
Plant Cells ◽  
Fluorescein Isothiocyanate ◽  
Kinetic Studies ◽  
Apparent Difference ◽  
Carrot Cells ◽  
Intracellular Compartments ◽  
Vacuolar System

The uptake of pure non-conjugated fluorescein isothiocyanate (FITC) and of the membraneimpermeant probe FITC—dextran into suspensioncultured carrot cells and protoplasts has been investigated. Commercial samples of a 70K (K=103Mr) FITC—dextran were shown to contain contaminant FITC and/or its degradation products, which were rapidly internalised into the vacuolar system of both cells and protoplasts. However, purified samples of the 70K FITC—dextran were taken up into the vacuoles of cells but not protoplasts after a lh incubation period. This apparent difference in the ability of cells and protoplasts to internalise FITC—dextrans was confirmed using samples of both commercial and purified 20K FITC—dextran as putative endocytotic probes. Both confocal and conventional fluorescence microscopy of FITC—treated cells have shown that FITC was internalised into similar intracellular compartments as was observed in cells treated with three-times purified 70K FITC—dextran. Thus, FITC was a useful fluorophore for rapidly labelling both the putative endocytotic compartments and the pleiomorphic vacuolar system of carrot cells. Kinetic studies indicated that FITC entered the cell by diffusion in the form of the neutral molecule. We have shown that treatment of cells or protoplasts with the drug Probenecid reversibly inhibited the uptake of FITC from the cytoplasm into the vacuole. In addition, the uptake of FITC into isolated vacuoles was enhanced in the presence of Mg-ATP

Stimulated release of fluorescently labeled IgE fragments that efficiently accumulate in secretory granules after endocytosis in RBL-2H3 mast cells

1998 ◽  
Vol 111 (16) ◽  
pp. 2385-2396 ◽  
Author(s):  
K. Xu ◽  
R.M. Williams ◽  
D. Holowka ◽  
B. Baird
Keyword(s):  
Mast Cells ◽  
Fluorescence Microscopy ◽  
High Efficiency ◽  
Immunoglobulin E ◽  
Secretory Granules ◽  
Dose Dependence ◽  
Secretory Vesicles ◽  
Release Process ◽  
Intracellular Compartments ◽  
Stimulation Of

Sensitization of RBL-2H3 mast cells with monomeric fluorescein-5-isothiocyanate (FITC)-labeled immunoglobulin E (IgE) results in slow but highly efficient accumulation of labeled IgE fragments in a pool of acidic peripheral vesicles that are visible by fluorescence microscopy after raising endosomal pH with ammonium chloride. Stimulation of cells containing these FITC-IgE fragments by aggregation of high affinity receptors for IgE (FcepsilonRI) or by Ca2+ ionophore and phorbol 12-myristate 13-acetate results in release of FITC fluorescence from the cells, which can be monitored continuously with a spectrofluorometer. The fluorescence release process corresponds to cellular degranulation: it is prevented under conditions that prevent stimulated beta-hexosaminidase release, and these two processes exhibit the same antigen dose-dependence and kinetics. Pulse-chase labeling reveals that aggregation of FITC-IgE bound to FcepsilonRI at the cell surface causes internalization and delivery to the regulated secretory vesicles with a high efficiency similar to monomeric IgE-FcepsilonRI, but more rapidly. Binding of Cy3-modified IgE to FcepsilonRI results in labeling of the same secretory vesicles as in FITC-IgE-sensitized cells, and these Cy3-labeled vesicles can be observed by fluorescence microscopy without neutralization of intracellular compartments. Simultaneous three-photon microscopy of serotonin fluorescence and two-photon microscopy of Cy3 fluorescence reveals that these Cy3-labeled vesicles coincide with serotonin-labeled secretory granules. After stimulation of the cells via aggregation of IgE-FcepsilonRI or addition of Ca2+ ionophore and phorbol 12-myristate 13-acetate, depletion of the Cy3 label from the intracellular vesicles is observed with confocal microscopy. These results provide strong evidence for the lysosomal nature of secretory granules in these cells. In addition, they provide the basis for a direct, real-time method for monitoring single cell degranulation.

Observations on the Aleurone Layer. II. Fluorescence Microscopy of the Aleurone-Sub-Aleurone Junction With Emphasis on Possible β-1,3-Glucan Deposits in Barley

1977 ◽  
Vol 4 (6) ◽  
pp. 917 ◽  
Author(s):  
RG Fulcher ◽  
G Setterfield ◽  
ME Mccully ◽  
PJ Wood
Keyword(s):  
Hordeum Vulgare ◽  
Fluorescence Microscopy ◽  
Cell Walls ◽  
Structural Organization ◽  
Periodate Oxidation ◽  
Aleurone Layer ◽  
Hordeum Vulgare L ◽  
Aleurone Cell ◽  
Tissue Sections ◽  
Primary Substrate

The structural organization of the cell walls at the aleurone-sub-aleurone junction has been examined by fluorescence microscopy in ungerminated seeds of two varieties of Hordeum vulgare L. (cultivars Himalaya and Vanier). The sub-aleurone cell walls that are immediately adjacent to the aleurone layer are considerably thicker than the remainder of endosperm walls and contain extensive deposits of aniline blue-positive material. The latter was not significantly affected by periodate oxidation and was removed from tissue sections only by β-1,3-glucanases or hot dimethyl sulphoxide. These deposits may represent the primary substrate for endo-β-1,3-glucanases secreted by the aleurone layer during germination.

Synchrotron-based X-ray Fluorescence Microscopy in Conjunction with Nanoindentation to Study Molecular-Scale Interactions of Phenol–Formaldehyde in Wood Cell Walls

2015 ◽  
Vol 7 (12) ◽  
pp. 6584-6589 ◽  
Author(s):  
Joseph E. Jakes ◽  
Christopher G. Hunt ◽  
Daniel J. Yelle ◽  
Linda Lorenz ◽  
Kolby Hirth ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Walls ◽  
Phenol Formaldehyde ◽  
X Ray ◽  
Scale Interactions ◽  
Molecular Scale

Proteomic response of the biological control fungus Trichoderma atroviride to growth on the cell walls of Rhizoctonia solani

2005 ◽  
Vol 47 (6) ◽  
pp. 381-388 ◽  
Author(s):  
Jasmine Grinyer ◽  
Sybille Hunt ◽  
Matthew McKay ◽  
Ben R. Herbert ◽  
Helena Nevalainen
Keyword(s):  
Biological Control ◽  
Rhizoctonia Solani ◽  
Cell Walls ◽  
Trichoderma Atroviride ◽  
Proteomic Response

scholarly journals A New View into the Silica Deposition Vesicles of Diatoms

2020 ◽  
Author(s):  
Christoph Heintze ◽  
Petr Formanek ◽  
Darius Pohl ◽  
Jannes Hauptstein ◽  
Bernd Rellinghaus ◽  
...  
Keyword(s):  
Cell Walls ◽  
High Spatial Resolution ◽  
Rapid Identification ◽  
Entire Volume ◽  
Microscopy Imaging ◽  
Silica Deposition ◽  
X Ray ◽  
Intracellular Compartments ◽  
Spatio Temporal ◽  
20 Nm

Abstract Diatoms are single-celled microalgae that produce silica-based cell walls with intricate nano- and micropatterns. Biogenesis of diatom biosilica is a bottom-up process that occurs in large intracellular compartments termed silica deposition vesicles (SDVs). Investigating the mechanism of silica morphogenesis has so far been severely limited by the lack of methods for imaging the entire volume of an SDV with high spatial resolution during all stages of development. Here we have developed a method that allows for rapid identification and electron microscopy imaging of many different, full sized SDVs that are in the process of producing biosilica valves. This enabled visualizing the development of characteristic morphological biosilica features with unprecedented spatio-temporal resolution. During early to mid-term development, valve SDVs contained ~ 20 nm sized particles that were primarily associated with the radially expanding rib-like biosilica structures. The results from electron dispersive X-ray analysis suggests that the immature biosilica patterns are silica-organic composites. This supports the hypothesis that silica morphogenesis is dependent on organic biomolecules inside the SDV lumen.

scholarly journals Candida utilis ATCC 9950 Cell Walls and β(1,3)/(1,6)-Glucan Preparations Produced Using Agro-Waste as a Mycotoxins Trap

Toxins ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 192 ◽  
Author(s):  
Anna Bzducha-Wróbel ◽  
Marcin Bryła ◽  
Iwona Gientka ◽  
Stanisław Błażejak ◽  
Monika Janowicz
Keyword(s):  
Cell Walls ◽  
Candida Utilis ◽  
Binding Capacity ◽  
Fumonisin B1 ◽  
Active Surface ◽  
Market Analysis ◽  
Feed Additives ◽  
Yeast Biomass ◽  
Food And Feed ◽  
Acidic Conditions

Mycotoxins are harmful contaminants of food and feed worldwide. Feed additives with the abilities to trap mycotoxins are considered substances which regulate toxin transfer from feed to tissue, reducing its absorption in animal digestive tract. Market analysis emphasizes growing interest of feed producers in mycotoxins binders obtained from yeast biomass. The aim of the study was to prescreen cell walls (CW) and β(1,3)/(1,6)-glucan (β-G) preparations isolated from Candida utilis ATCC 9950 cultivated on waste potato juice water with glycerol as adsorbents for aflatoxin B1 (AFB1), zearalenone (ZEN), ochratoxin A (OTA), deoxynivalenol (DON), nivalenol (NIV), T-2 toxin (T-2) and fumonisin B1 (FB1). The adsorption was studied in single concentration tests at pH 3.0 and 6.0 in the presence of 1% of the adsorbent and 500 ng/mL of individual toxin. Evaluated CW and β-G preparations had the potential to bind ZEN, OTA and AFB1 rather than DON, NIV, T-2 toxin and FB1. The highest percentage of adsorption (about 83%), adsorption capacity (approx. 41 µg/ g preparation) and distribution coefficient (458.7mL/g) was found for zearalenone when CW preparation was used under acidic conditions. Higher protein content in CW and smaller particles sizes of the formulation could influence more efficient binding of ZEN, OTA, DON and T-2 toxin at appropriate pH compared to purified β-G. Obtained results show the possibility to transform the waste potato juice water into valuable Candida utilis ATCC 9950 preparation with mycotoxins adsorption properties. Further research is important to improve the binding capacity of studied preparations by increasing the active surface of adsorption.

scholarly journals Stiffness tomography of eukaryotic intracellular compartments by atomic force microscopy

Nanoscale ◽  
2019 ◽  
Vol 11 (21) ◽  
pp. 10320-10328 ◽  
Author(s):  
Sébastien Janel ◽  
Michka Popoff ◽  
Nicolas Barois ◽  
Elisabeth Werkmeister ◽  
Séverine Divoux ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Atomic Force Microscopy ◽  
Fluorescence Microscopy ◽  
Force Microscopy ◽  
Atomic Force ◽  
Ultrastructural Characterization ◽  
Intracellular Compartments

After identification by fluorescence microscopy, intracellular compartments are analyzed by stiffness tomography using atomic force microscopy, before further processing for ultrastructural characterization by electron microscopy.

scholarly journals Endo-β-1,3-glucanase (GH16 Family) from Trichoderma harzianum Participates in Cell Wall Biogenesis but Is Not Essential for Antagonism Against Plant Pathogens

Biomolecules ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 781 ◽  
Author(s):  
Marcela Suriani Ribeiro ◽  
Renato Graciano de Paula ◽  
Aline Raquel Voltan ◽  
Raphaela Georg de Castro ◽  
Cláudia Batista Carraro ◽  
...  
Keyword(s):  
Cell Wall ◽  
Trichoderma Harzianum ◽  
Cell Walls ◽  
Plant Pathogens ◽  
Pcr Analysis ◽  
Glycosyl Hydrolases ◽  
Lytic Enzymes ◽  
Trichoderma Spp ◽  
Compensatory Response ◽  
Trichoderma Species

Trichoderma species are known for their ability to produce lytic enzymes, such as exoglucanases, endoglucanases, chitinases, and proteases, which play important roles in cell wall degradation of phytopathogens. β-glucanases play crucial roles in the morphogenetic-morphological process during the development and differentiation processes in Trichoderma species, which have β-glucans as the primary components of their cell walls. Despite the importance of glucanases in the mycoparasitism of Trichoderma spp., only a few functional analysis studies have been conducted on glucanases. In the present study, we used a functional genomics approach to investigate the functional role of the gluc31 gene, which encodes an endo-β-1,3-glucanase belonging to the GH16 family in Trichoderma harzianum ALL42. We demonstrated that the absence of the gluc31 gene did not affect the in vivo mycoparasitism ability of mutant T. harzianum ALL42; however, gluc31 evidently influenced cell wall organization. Polymer measurements and fluorescence microscopy analyses indicated that the lack of the gluc31 gene induced a compensatory response by increasing the production of chitin and glucan polymers on the cell walls of the mutant hyphae. The mutant strain became more resistant to the fungicide benomyl compared to the parental strain. Furthermore, qRT-PCR analysis showed that the absence of gluc31 in T. harzianum resulted in the differential expression of other glycosyl hydrolases belonging to the GH16 family, because of functional redundancy among the glucanases.

scholarly journals Identification of a New Biocontrol Gene in Trichoderma atroviride: The Role of an ABC Transporter Membrane Pump in the Interaction with Different Plant-Pathogenic Fungi

2009 ◽  
Vol 22 (3) ◽  
pp. 291-301 ◽  
Author(s):  
Michelina Ruocco ◽  
Stefania Lanzuise ◽  
Francesco Vinale ◽  
Roberta Marra ◽  
David Turrà ◽  
...  
Keyword(s):  
Pathogenic Fungi ◽  
Pythium Ultimum ◽  
Resistance Mechanisms ◽  
Trichoderma Atroviride ◽  
Related Factors ◽  
Trichoderma Spp ◽  
Membrane Pump ◽  
Synthetic Chemicals ◽  
Inhibitory Compounds

Successful biocontrol interactions often require that the beneficial microbes involved are resistant or tolerant to a variety of toxicants, including antibiotics produced by themselves or phytopathogens, plant antimicrobial compounds, and synthetic chemicals or contaminants. The ability of Trichoderma spp., the most widely applied biocontrol fungi, to withstand different chemical stresses, including those associated with mycoparasitism, is well known. In this work, we identified an ATP-binding cassette transporter cell membrane pump as an important component of the above indicated resistance mechanisms that appears to be supported by an extensive and powerful cell detoxification system. The encoding gene, named Taabc2, was cloned from a strain of Trichoderma atroviride and characterized. Its expression was found to be upregulated in the presence of pathogen-secreted metabolites, specific mycotoxins and some fungicides, and in conditions that stimulate the production in Trichoderma spp. of antagonism-related factors (toxins and enzymes). The key role of this gene in antagonism and biocontrol was demonstrated by the characterization of the obtained deletion mutants. They suffered an increased susceptibility to inhibitory compounds either secreted by pathogenic fungi or possibly produced by the biocontrol microbe itself and lost, partially or entirely, the ability to protect tomato plants from Pythium ultimum and Rhizoctonia solani attack.

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