In plantamonitoring of the activity of two constitutive promoters, CaMV 35S and TR2′, in developing feeding cells induced byGlobodera rostochiensisusing green fluorescent protein in combination with confocal laser scanning microscopy

1998 ◽  
Vol 52 (4) ◽  
pp. 275-284 ◽  
Author(s):  
A Goverse ◽  
J Biesheuvel ◽  
G.-J Wijers ◽  
F.J Gommers ◽  
J Bakker ◽  
...  
2014 ◽  
Vol 20 (3) ◽  
pp. 912-915 ◽  
Author(s):  
Brandon W. Peterson ◽  
Henk J. Busscher ◽  
Prashant K. Sharma ◽  
Henny C. van der Mei

AbstractBacterial biofilms relieve themselves from external stresses through internal rearrangement, as mathematically modeled in many studies, but never microscopically visualized for their underlying microbiological processes. The aim of this study was to visualize rearrangement processes occurring in mechanically deformed biofilms using confocal-laser-scanning-microscopy after SYTO9 (green-fluorescent) and calcofluor-white (blue-fluorescent) staining to visualize bacteria and extracellular-polymeric matrix substances, respectively. We apply 20% uniaxial deformation toPseudomonas aeruginosabiofilms and fix deformed biofilms prior to staining, after allowing different time-periods for relaxation. Two isogenicP. aeruginosastrains with different abilities to produce extracellular polymeric substances (EPS) were used. By confocal-laser-scanning-microscopy all biofilms showed intensity distributions for fluorescence from which rearrangement of EPS and bacteria in deformed biofilms were derived. For theP. aeruginosastrain producing EPS, bacteria could not find new, stable positions within 100 s after deformation, while EPS moved toward deeper layers within 20 s. Bacterial rearrangement was not seen inP. aeruginosabiofilms deficient in production of EPS. Thus, EPS is required to stimulate bacterial rearrangement in mechanically deformed biofilms within the time-scale of our experiments, and the mere presence of water is insufficient to induce bacterial movement, likely due to its looser association with the bacteria.


2002 ◽  
Vol 15 (2) ◽  
pp. 172-179 ◽  
Author(s):  
Anastasia L. Lagopodi ◽  
Arthur F. J. Ram ◽  
Gerda E. M. Lamers ◽  
Peter J. Punt ◽  
Cees A. M. J. J. Van den Hondel ◽  
...  

The fungus Fusarium oxysporum f. sp. radicis-lycopersici is the causal agent of tomato foot and root rot disease. The green fluorescent protein (GFP) was used to mark this fungus in order to visualize and analyze the colonization and infection processes in vivo. Transformation of F. oxysporum f. sp. radicis-lycopersici was very efficient and gfp expression was stable for at least nine subcultures. Microscopic analysis of the transformants revealed homogeneity of the fluorescent signal, which was clearly visible in the hyphae as well as in the chlamydospores and conidia. To our knowledge, this is the first report in which this is shown. The transformation did not affect the pathogenicity. Using confocal laser scanning microscopy, colonization, infection, and disease development on tomato roots were visualized in detail and several new aspects of these processes were observed, such as (i) the complete colonization pattern of the tomato root system; (ii) the very first steps of contact between the fungus and the host, which takes place at the root hair zone by mingling and by the attachment of hyphae to the root hairs; (iii) the preferential colonization sites on the root surface, which are the grooves along the junctions of the epidermal cells; and (iv) the absence of specific infection sites, such as sites of emergence of secondary roots, root tips, or wounded tissue, and the absence of specific infection structures, such as appressoria. The results of this work prove that the use of GFP as a marker for F. oxysporum f. sp. radicis-lycopersici is a convenient, fast, and effective approach for studying plant-fungus interactions.


2012 ◽  
Vol 11 (3) ◽  
pp. 669-674 ◽  
Author(s):  
Szabolcs Szilveszter ◽  
Botond Raduly ◽  
Szilard Bucs ◽  
Beata Abraham ◽  
Szabolcs Lanyi ◽  
...  

2021 ◽  
Vol 11 (8) ◽  
pp. 3403
Author(s):  
Shlomo Elbahary ◽  
Sohad Haj Yahya ◽  
Cemre Koç ◽  
Hagay Shemesh ◽  
Eyal Rosen ◽  
...  

Following furcal perforation, bacteria may colonize the defect and cause inflammation and periodontal destruction. This study used confocal laser scanning microscopy (CLSM) to evaluate Enterococcus faecalis colonization and proliferation in furcal perforations repaired with different materials. Furcal perforations created in 55 extracted human mandibular molars were repaired using either MTA-Angelus, Endocem, or Biodentine and coronally subjected to E. faecalis suspension for 21 days. The specimens were then stained using a LIVE/DEAD Viability Kit and visualized by CLSM. The minimum and maximum depths of bacterial penetration into the dentinal tubules were 159 and 1790 μM, respectively, with a mean of 713 μM. There were significantly more live than dead bacteria inside the dentinal tubules (p = 0.0023) in all groups, and all three repair materials exhibited a similarly sized stained area (p = 0.083). However, there were significant differences in the numbers of dead bacteria at the circumference of the perforation defect (p = 0.0041), with a significantly higher ratio of live to dead bacteria in the MTA-Angelus group (p = 0.001). Following perforation repair, bacteria may colonize the interface between the repair material and dentin and may penetrate through the dentinal tubules. The type of repair material has a significant effect on the viability of the colonizing bacteria.


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