Protoplast Formation from Submerged Mycelium and from Spore Germinants of Streptomyces coelicolor

1985 ◽  
Vol 38 (1) ◽  
pp. 175 ◽  
Author(s):  
I Stevenson

Stages in the formation of protoplasts from S. coelicolor strain A3(2) have been studied by transmission electron microscopy. Protoplasts liberated from submerged mycelial growth were variable in size and were released when digestion of the cell wall by lysozyme had completely or almost completely taken place. Protoplasts did not fully adopt the typical rounded shape until after release. A single region of cytoplasm gave rise to more than one protoplast unit. Protoplasts released from spore germinants escaped from the tip of the germ tube, which was the region of the cell wall most susceptible to digestion. Protoplasts derived from spore germinants were more consistent in size and rounded up more rapidly. If a cross-wall had formed in a germinant then it gave rise to separate protoplasts from each cellular. compartment. Protoplasts of either type contained a single DNA region. These studies give an indication of the cellular organization of a streptomycete colony, which can be visualized as a multinucleated assemblage of cellular units in a common cytoplasm. The assembly of units separates into a number of protoplasts on digestion of the cell wall.

2006 ◽  
Vol 87 (1) ◽  
pp. 17-27 ◽  
Author(s):  
Guillemond B. Ouellette ◽  
Mohamed Cherif ◽  
Marie Simard

Abstract Various cell reactions occurred in staghorn sumac plants inoculated with Fusarium oxysporum f. sp. callistephi. Light and transmission electron microscopy observations and results of cytochemical tests showed: 1) increased laticifers and latex production in the phloem; 2) tylosis formation; 3) host cell wall modifications, including appositions or other cell wall thickenings; and 4) unusual cross wall formation in some cells, and cell hypertrophy and hyperplasia. Tylosis walls labelled for pectin and cellulose and many displayed inner suberin-like layers. These layers were also noted in cells of the medullary sheath and in many cells with dense content and thickened walls in the barrier zones that had formed. These zones also contained fibres with newly-formed gelatinous-like layers. In the vicinity of these cells, host cell walls were frequently altered, associated with opaque matter. Many small particles present in chains also occurred in some of these cells, which contained only remnants of host cytoplasm. Light microscopy observations showed that pronounced tissue proliferation and aberrant cells occurred in the outer xylem in the infected plants. Unusual neoplasmic tissue also formed from cells surrounding the pith and medullary sheath, and it spanned directly across the pre-existing xylem tissue and burst as large mounds on the stems.


1992 ◽  
Vol 55 (9) ◽  
pp. 687-696 ◽  
Author(s):  
SOUZAN E. EL-KEST ◽  
ELMER H. MARTH

Unfrozen cells of Listeria monocytogenes typically contained no preplasma space exterior to the plasma membrane (PM) when viewed by transmission electron microscopy. Cells of L. monocytogenes strains Scott A, V7, and California (CA), after freezing and frozen storage, exhibited one or more of the following when viewed with transmission electron microscopy: (a) retraction of cytoplasm and infolding of the PM to form mesosomes, (b) extra-and intracellular rupture of the cell wall (CW), (c) formation of intracellular “bubbles,” and (d) damage to the CW and PM that could have resulted from autolysin activity. Type and degree of effect depended on frozen storage time and strain of L. monocytogenes. Lysozyme treatment of unfrozen or frozen/stored (19 d)/thawed cells of strains Scott A, V7, and CA resulted in protoplast formation and damage to the CW. Three stages of protoplast formation were observed when cells of strain CA were frozen, stored 2 weeks, thawed, and treated with lysozyme. More damage to the CW and PM occurred when frozen storage time was extended for up to 6 weeks before treatment with lysozyme. Lipase and lysozyme treatment of unfrozen or frozen/stored (19 d)/thawed cells of strain Scott A resulted in protoplast formation with some damage to the PM and irregularity in shape of cells. Damage to the PM increased with increasing frozen storage time for up to 6 weeks. Some cells of strain CA resisted freezing, frozen storage for 6 weeks, thawing, and treatment with lipase and lysozyme.


1997 ◽  
Vol 161 ◽  
pp. 491-504 ◽  
Author(s):  
Frances Westall

AbstractThe oldest cell-like structures on Earth are preserved in silicified lagoonal, shallow sea or hydrothermal sediments, such as some Archean formations in Western Australia and South Africa. Previous studies concentrated on the search for organic fossils in Archean rocks. Observations of silicified bacteria (as silica minerals) are scarce for both the Precambrian and the Phanerozoic, but reports of mineral bacteria finds, in general, are increasing. The problems associated with the identification of authentic fossil bacteria and, if possible, closer identification of bacteria type can, in part, be overcome by experimental fossilisation studies. These have shown that not all bacteria fossilise in the same way and, indeed, some seem to be very resistent to fossilisation. This paper deals with a transmission electron microscope investigation of the silicification of four species of bacteria commonly found in the environment. The Gram positiveBacillus laterosporusand its spore produced a robust, durable crust upon silicification, whereas the Gram negativePseudomonas fluorescens, Ps. vesicularis, andPs. acidovoranspresented delicately preserved walls. The greater amount of peptidoglycan, containing abundant metal cation binding sites, in the cell wall of the Gram positive bacterium, probably accounts for the difference in the mode of fossilisation. The Gram positive bacteria are, therefore, probably most likely to be preserved in the terrestrial and extraterrestrial rock record.


Holzforschung ◽  
2005 ◽  
Vol 59 (6) ◽  
pp. 675-680 ◽  
Author(s):  
Jonas Brändström ◽  
Jean-Paul Joseleau ◽  
Alain Cochaux ◽  
Nathalie Giraud-Telme ◽  
Katia Ruel

Abstract Transmission electron microscopy was used to investigate the ultrastructure of recycled pulp fibers originating from a household collection plant and intended for the production of packaging paper. Three recovered paper grades and recycling processes, including pulping, screening, cleaning and refining, were assessed with emphasis on surface and internal fibrillation as well as xylan localization. Results showed a large heterogeneity with respect to fiber ultrastructure within and between the grades. Screening and cleaning steps had no detectable effects, but refining clearly increased cell-wall delamination and surface fibrillation. Immunolabeling of xylans showed that they were distributed rather evenly across the cell walls. They were also present on fines. Two different mechanisms for fiber delamination and surface fibrillation were found, one which implies that internal and external fibrillation take place simultaneously across the cell wall, and another which implies successive peeling of layers or sub-layers from the outside towards the inside. It is suggested that recycled fibers of chemical pulp origin undergo the former mechanism and recycled fibers that contain lignin binding the cell wall matrix give rise to the latter peeling mechanism. Because several recycled fibers were severely delaminated and almost fractured, we suggest that to produce a good packaging paper, it is important that recycled pulp should contain a significant proportion of fibers with high intrinsic strength.


2013 ◽  
Vol 57 (9) ◽  
pp. 4470-4480 ◽  
Author(s):  
Min Jung Kwun ◽  
Gabriela Novotna ◽  
Andrew R. Hesketh ◽  
Lionel Hill ◽  
Hee-Jeon Hong

ABSTRACTVanRS two-component regulatory systems are key elements required for the transcriptional activation of inducible vancomycin resistance genes in bacteria, but the precise nature of the ligand signal that activates these systems has remained undefined. Using the resistance system inStreptomyces coelicoloras a model, we have undertaken a series ofin vivostudies which indicate that the VanS sensor kinase in VanB-type resistance systems is activated by vancomycin in complex with thed-alanyl-d-alanine (d-Ala-d-Ala) termini of cell wall peptidoglycan (PG) precursors. Complementation of an essentiald-Ala-d-Ala ligase activity by constitutive expression ofvanAencoding a bifunctionald-Ala-d-Ala andd-alanyl-d-lactate (d-Ala-d-Lac) ligase activity allowed construction of strains that synthesized variable amounts of PG precursors containingd-Ala-d-Ala. Assays quantifying the expression of genes under VanRS control showed that the response to vancomycin in these strains correlated with the abundance ofd-Ala-d-Ala-containing PG precursors; strains producing a lower proportion of PG precursors terminating ind-Ala-d-Ala consistently exhibited a lower response to vancomycin. Pretreatment of wild-type cells with vancomycin or teicoplanin to saturate and mask thed-Ala-d-Ala binding sites in nascent PG also blocked the transcriptional response to subsequent vancomycin exposure, and desleucyl vancomycin, a vancomycin analogue incapable of interacting withd-Ala-d-Ala residues, failed to inducevangene expression. Activation of resistance by a vancomycin–d-Ala-d-Ala PG complex predicts a limit to the proportion of PG that can be derived from precursors terminating ind-Ala-d-Lac, a restriction also enforced by the bifunctional activity of the VanA ligase.


1983 ◽  
Vol 61 (12) ◽  
pp. 3444-3453 ◽  
Author(s):  
R. N. Trigiano ◽  
C. G. Van Dyke ◽  
H. W. Spurr Jr.

The development of haustoria in tobacco by the blue-mold fungus Peronospora tabacina was examined using light, scanning, and transmission electron microscopy. Electron-lucent, callose-like appositions were observed between the host plasmalemma and the host mesophyll cell wall prior to haustorial penetration. An electron-opaque penetration matrix was present between the apposition and the host cell wall. The intercellular hyphal wall consisted of two layers which differed in staining quality. The haustorial wall was also two layered, but was primarily composed of and continuous with the inner wall layer of the intercellular hypha. Haustoria were either finger-like or branched and were encased with callose-like material. Most encasements were thickened at the proximal regions of haustoria but were thinner along the distal portions. Vesicles were present in host cytoplasm and were occasionally attached to the invaginated host plasmalemma. These vesicles might contribute to the deposition of the encasement material. The encasement stained positively for callose using aniline blue; calcofluor and toluidine blue O tests for cellulose were inconclusive, and lignin was not detected using toluidine blue O or phloroglucinol–HCl.


Biomolecules ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 703
Author(s):  
Suriya Rehman ◽  
Munirah A. Almessiere ◽  
Ebtesam A. Al-Suhaimi ◽  
Mehwish Hussain ◽  
Maha Yousuf Bari ◽  
...  

In the present study, biocompatible manganese nanoparticles have been linked with zinc and iron molecules to prepare different derivatives of Mn0.5Zn0.5ErxYxFe2−2xO4 NPs (x = 0.02, 0.04, 0.06, 0.08, 0.10), using an ultrasonication approach. The structure, surface morphology, and chemical compositions of Mn0.5Zn0.5ErxYxFe2−2xO4 NPs were elucidated by X-ray diffractometer (XRD), High-resolution transmission electron microscopy (HR-TEM), scanning electron microscope (SEM), and Energy Dispersive X-Ray Analysis (EDX) techniques. The bioactivity of Mn0.5Zn0.5ErxYxFe2−2xO4 NPs on normal (HEK-293) and (HCT-116) colon cancer cell line was evaluated. The Mn0.5Zn0.5ErxYxFe2−2xO4 NPs treatment post 48 h resulted in a significant reduction in cells (via MTT assay, having an IC50 value between 0.88 µg/mL and 2.40 µg/mL). The specificity of Mn0.5Zn0.5ErxYxFe2−2xO4 NPs were studied by treating them on normal cells line (HEK-293). The results showed that Mn0.5Zn0.5ErxYxFe2−2xO4 NPs did not incur any effect on HEK-293, which suggests that Mn0.5Zn0.5ErxYxFe2−2xO4 NPs selectively targeted the colon cancerous cells. Using Candida albicans, antifungal activity was also studied by evaluating minimum inhibitory/fungicidal concentration (MIC/MFC) and the effect of nanomaterial on the germ tube formation, which exhibited that NPs significantly inhibited the growth and germ tube formation. The obtained results hold the potential to design nanoparticles that lead to efficient bioactivity.


1992 ◽  
Vol 102 (3) ◽  
pp. 611-627 ◽  
Author(s):  
I. BRENT HEATH ◽  
RUTH L. HAROLD

Very similar changing patterns of actin are described with rhodamine-phalloidin labelling during the zoosporic life cycle of the oomycetes, Saprolegnia ferax and Achlya bisexualis. By comparing the changes with previously described ultrastructural and functional changes, we show that actin functions in numerous previously unrecognized processes. Most spectacularly, the directed vesicle expansions of the cytokinetic system involve newly formed actin which outlines the developing zoospores. Disruption of this actin with cytochalasins leads to abnormal cleavage as witnessed by the formation of enlarged and irregular cysts. Prior to cytokinesis, two new types of organelle are synthesized and one, known as K bodies, clusters around the nuclei. These organdies are actin-rich during development and clustering, consistent with actin functioning in their positioning. In the zoospores, actin is concentrated around the water expulsion vacuoles, indicating that they are contractile, and permeates the cytoplasm, probably with a skeletal role. This concept is supported by the first demonstration of actin specifically associated with a microtubular root in the secondary zoospore. Upon encystment there is a dramatic increase in stained actin in the form of peripheral plaques associated with the newly synthesized cell wall. When the cysts germinate, a fibrillar actin cap, comparable to that previously described in hyphal tips, forms in the germ tube apex, but only after cell wall softening to permit germ tube protrusion. This sequence is consistent with the actin cap modulating turgor-driven expansion of the tip as previously discussed for hyphae. In addition to disrupting cleavage-associated actin, cytochalasins show developmental stage, dose and drug (CE≥CD≥CB) specific effects on zoosporulation-related actin, which indicates that, contrary to previous suggestions, rhodamine-phalloidin staining is a useful indicator of actin behaviour in response to cytochalasins. These responses include differential effects on adjoining actin arrays, some of which are transient in the continued presence of the drugs, indicating a mechanism of drug adaptation.


Plant Disease ◽  
2021 ◽  
Author(s):  
Xue Yang ◽  
Chun-Yan Gu ◽  
Yang Bai ◽  
Jia-Zhi Sun ◽  
Hao-Yu Zang ◽  
...  

Pomegranate crown rot caused by Coniellagranati is one of the most severe diseases of pomegranate. To date, no fungicides have been registered for controlling this disease in China. Pyraclostrobin, belonging to strobilurin fungicides, has a broad spectrum of activity against many phytopathogens. In this study, based on the mycelial growth and conidial germination inhibition methods, we investigated the biological activity of pyraclostrobin against C. granati at the presence of 50 μg/mL SHAM using 80 isolates collected from different orchards in China during 2012-2018. The EC50 (50% effective concentration) values ranged from 0.040-0.613 μg/mL for mycelial growth and 0.013-0.110 μg/mL for conidium germination, respectively. Treated with pyraclostrobin, the hyphae morphology changed and conidial production of C. granati decreased significantly. The result of transmission electron microscope showed that treatment of pyraclostrobin could make the cell wall thinner, and lead to ruptured cell membrane and formation of intracellular organelle autophagosomes. The pyraclostrobin showed good protective and curative activities against C. granati on detached pomegranate fruits. In field trials, pyraclostrobin showed excellent control efficacy against this disease in which the treatment of 25% pyraclostrobin EC 1000× provided 92.25% and 92.58% control efficacy in 2019 and 2020, respectively, significantly higher than that of other treatments. Therefore, pyraclostrobin could be a candidate fungicide for the control of pomegranate crown rot.


Sign in / Sign up

Export Citation Format

Share Document