scholarly journals Endophytic Bacillus cereus Effectively Controls Meloidogyne incognita on Tomato Plants Through Rapid Rhizosphere Occupation and Repellent Action

Plant Disease ◽  
2017 ◽  
Vol 101 (3) ◽  
pp. 448-455 ◽  
Author(s):  
Hai-Jing Hu ◽  
Ya-Li Chen ◽  
Yu-Fang Wang ◽  
Yun-Yun Tang ◽  
Shuang-Lin Chen ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Meloidogyne Incognita ◽  
Endophytic Bacteria ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Egg Mass ◽  
Root Knot Nematodes ◽  
Tomato Roots ◽  
Repellent Action

Root-knot nematodes (Meloidogyne spp.), which cause severe global agricultural losses, can establish a special niche in the root vascular cylinder of crops, making them difficult to control. Endophytic bacteria have great potential as biocontrol organisms against Meloidogyne incognita. Three endophytic bacteria were isolated from plant tissues and showed high nematicidal activity against M. incognita second-stage juveniles (J2) in vitro. The gyrB gene sequence amplification results indicated that the three isolates were Bacillus cereus BCM2, B. cereus SZ5, and B. altitudinis CCM7. The isolates colonized tomato roots rapidly and stably during the colonization dynamic experiment. Three pot experiments were designed to determine the potential of three endophytic bacterial isolates on control of root-knot nematodes. The results showed that the preinoculated B. cereus BCM2 experiment significantly reduced gall and egg mass indexes. The inhibition ratio of gall and egg mass was up to 81.2 and 75.6% on tomato roots and significantly enhanced shoot length and fresh weight. The other two experiments with inoculated endophytic bacteria and M. incognita at the same time or after morbidity had lower inhibition ratios compared with the preinoculated endophytic bacteria experiment. The confocal laser-scanning microscopy method was used to further study the possible mechanism of endophytic bacteria in the biocontrol process. The results showed the localization pattern of the endophytic bacteria B. cereus BCM2-(str′)-pBCgfp-1 in tomato root tissues. Root tissue colonized by endophytic bacteria repelled M. incognita J2 infection compared with the untreated control in a repellence experiment. We isolated an endophytic B. cereus strain that stably colonized tomato and controlled M. incognita effectively. This strain has potential for plant growth promotion, successful ecological niche occupation, and M. incognita J2 repellent action induction. It plays an important role in endophytic bacteria against root-knot nematodes.

Biocontrol efficacy of Bacillus cereus strain Bc-cm103 against Meloidogyne incognita

Plant Disease ◽  
2021 ◽  
Author(s):  
Nan Yin ◽  
JianLong zhao ◽  
Rui Liu ◽  
Yan Li ◽  
Jian Ling ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Meloidogyne Incognita ◽  
Laser Scanning ◽  
Fermentation Broth ◽  
Laser Scanning Microscopy ◽  
Economic Losses ◽  
Confocal Laser ◽  
Hatching Rate ◽  
Egg Hatching ◽  
Root Knot Nematodes

Root-knot nematodes (Meloidogyne spp.) are soil-borne pathogens that infect vegetable crops and cause major economic losses worldwide annually. Therefore, there is an urgent need for novel nematicides or biological control agents to reduce the damage caused by root-knot nematodes. In this study, we tested efficacy of the Bacillus cereus strain Bc-cm103, isolated from the rhizoplane of Cucumis metuliferus, against Meloidogyne incognita. The strain Bc-cm103 fermentation broth caused 100% mortality of the nematode second-stage juveniles (J2s) within 12 h and decreased the egg hatching rate by 40.06% within 72 h compared with sterile water. Confocal laser scanning microscopy revealed that strain Bc-cm103 formed a biofilm on cucumber (Cucumis sativus) roots, which protected the roots from the infection of M. incognita. Additionally, strain Bc-cm103 activated the defense-responsive genes PR1, PR2, LOX1, and CTR1 in cucumber. Furthermore, strain Bc-cm103 significantly reduced the appearance of root galls in pot, split-root and field tests. These results indicated that the B. cereus strain Bc-cm103 had a strong suppressive effect on M. incognita and therefore could be used as a potential biocontrol agent against this pathogen.

scholarly journals Effects of the Endophytic Bacteria Bacillus cereus BCM2 on Tomato Root Exudates and Meloidogyne incognita Infection

Plant Disease ◽  
2019 ◽  
Vol 103 (7) ◽  
pp. 1551-1558 ◽  
Author(s):  
Xia Li ◽  
Hai-Jing Hu ◽  
Jing-Yu Li ◽  
Cong Wang ◽  
Shuang-Lin Chen ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Root Exudates ◽  
Meloidogyne Incognita ◽  
Endophytic Bacteria ◽  
Root Exudate ◽  
Tomato Root ◽  
Second Stage ◽  
Tomato Roots ◽  
Meloidogyne Spp ◽  
Root Tissues

Root-knot nematodes (Meloidogyne spp.) cause serious crop losses worldwide. The colonization of tomato roots by endophytic bacteria Bacillus cereus BCM2 can greatly reduce Meloidogyne incognita damage, and tomato roots carrying BCM2 were repellent to M. incognita second-stage juveniles (J2). Here, the effects of BCM2 colonization on the composition of tomato root exudates was evaluated and potential mechanisms for BCM2-mediated M. incognita control explored using a linked twin-pot assay and GC-MS. On water agar plates, J2 preferentially avoided filter paper treated with tomato root exudates (organic phase only) from plants inoculated with BCM2, visiting these 67.1% less than controls. In a linked twin-pot assay, BCM2 treatment resulted in a 42.0% reduction in the number of nematodes in the soil, a 43.3% reduction in the number of galls and a 47.7% decrease in the density of M. incognita in root tissues. Analysis of root exudate composition revealed that BCM2 inoculation increased the number of components in exudates. Among these, 2,4-di-tert-butylphenol, 3,3-dimethyloctane, and n-tridecane secretions markedly increased. In repellency trials on water agar plates, J2 avoided 2,4-di-tert-butylphenol, n-tridecane, and 3,3-dimethyloctane at concentrations of 4 mmol/liter. In a linked twin-pot assay, inoculation with 2,4-di-tert-butylphenol or 3,3-dimethyloctane reduced the number of nematodes in the soil (by 54.9 and 70.6%, respectively), the number of galls (by 53.7 and 52.4%), and the number of M. incognita in root tissues (by 67.5 and 36.3%). BCM2 colonization in tomato roots affected the composition of root exudates, increasing the secretion of substances that appear to be repellent, thus decreasing M. incognita J2 infection of roots.

scholarly journals Colonization of Tomato Root by Pathogenic and Nonpathogenic Fusarium oxysporum Strains Inoculated Together and Separately into the Soil

2006 ◽  
Vol 72 (2) ◽  
pp. 1523-1531 ◽  
Author(s):  
Chantal Olivain ◽  
Claude Humbert ◽  
Jarmila Nahalkova ◽  
Jamshid Fatehi ◽  
Floriane L'Haridon ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Laser Scanning ◽  
Root Surface ◽  
Laser Scanning Microscopy ◽  
Pathogenic Strain ◽  
Confocal Laser ◽  
Fungal Colonization ◽  
Tomato Root ◽  
Tomato Roots ◽  
Nonpathogenic Strain

ABSTRACT In soil, fungal colonization of plant roots has been traditionally studied by indirect methods such as microbial isolation that do not enable direct observation of infection sites or of interactions between fungal pathogens and their antagonists. Confocal laser scanning microscopy was used to visualize the colonization of tomato roots in heat-treated soil and to observe the interactions between a nonpathogenic strain, Fo47, and a pathogenic strain, Fol8, inoculated onto tomato roots in soil. When inoculated separately, both fungi colonized the entire root surface, with the exception of the apical zone. When both strains were introduced together, they both colonized the root surface and were observed at the same locations. When Fo47 was introduced at a higher concentration than Fol8, it colonized much of the root surface, but hyphae of Fol8 could still be observed at the same location on the root. There was no exclusion of the pathogenic strain by the presence of the nonpathogenic strain. These results are not consistent with the hypothesis that specific infection sites exist on the root for Fusarium oxysporum and instead support the hypothesis that competition occurs for nutrients rather than for infection sites.

scholarly journals Visualization of Interactions Between a Pathogenic and a Beneficial Fusarium Strain During Biocontrol of Tomato Foot and Root Rot

2005 ◽  
Vol 18 (7) ◽  
pp. 710-721 ◽  
Author(s):  
Annouschka Bolwerk ◽  
Anastasia L. Lagopodi ◽  
Ben J. J. Lugtenberg ◽  
Guido V. Bloemberg
Keyword(s):  
Root Rot ◽  
Laser Scanning ◽  
Root Colonization ◽  
Pathogenic Fungus ◽  
Laser Scanning Microscopy ◽  
Systemic Resistance ◽  
Confocal Laser ◽  
Tomato Root ◽  
Tomato Roots

The soilborne fungus Fusarium oxysporum f. sp. radicislycopersici causes tomato foot and root rot (TFRR), which can be controlled by the addition of the nonpathogenic fungus F. oxysporum Fo47 to the soil. To improve our understanding of the interactions between the two Fusarium strains on tomato roots during biocontrol, the fungi were labeled using different autofluorescent proteins as markers and subsequently visualized using confocal laser scanning microscopy. The results were as follows. i) An at least 50- fold excess of Fo47over F. oxysporum f. sp. radicis-lycopersici was required to obtain control of TFRR. ii) When seedlings were planted in sand infested with spores of a single fungus, Fo47 hyphae attached to the root earlier than those of F. oxysporum f. sp. radicis-lycopersici. iii) Subsequent root colonization by F. oxysporum f. sp. radicis-lycopersici was faster and to a larger extent than that by Fo47. iv) Under disease-controlling conditions, colonization of tomato roots by the pathogenic fungus was significantly reduced. v) When the inoculum concentration of Fo47 was increased, root colonization by the pathogen was arrested at the stage of initial attachment to the root. vi) The percentage of spores of Fo47 that germinates in tomato root exudate in vitro is higher than that of the pathogen F. oxysporum f. sp. radicis-lycopersici. Based on these results, the mechanisms by which Fo47 controls TFRR are discussed in terms of i) rate of spore germination and competition for nutrients before the two fungi reach the rhizoplane; ii) competition for initial sites of attachment, intercellular junctions, and nutrients on the tomato root surface; and iii) inducing systemic resistance.

3-D imaging of cells using a confocal laser scanning microscope and digital image processing

1988 ◽  
Vol 46 ◽  
pp. 96-97
Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Laser Scanning Microscope ◽  
Biochemical Processes ◽  
Adjacent Structures

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

Use of confocal laser scanning microscopy and a computer model to understand ink cavitation and filamentation

TAPPI Journal ◽  
2010 ◽  
Vol 9 (10) ◽  
pp. 7-15
Author(s):  
HANNA KOIVULA ◽  
DOUGLAS BOUSFIELD ◽  
MARTTI TOIVAKKA
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Print Quality ◽  
Length Scale ◽  
Offset Printing ◽  
Significant Difference ◽  
Printing Speed

In the offset printing process, ink film splitting has an important impact on formation of ink filaments. The filament size and its distribution influence the leveling of ink and hence affect ink setting and the print quality. However, ink filaments are difficult to image due to their short lifetime and fine length scale. Due to this difficulty, limited work has been reported on the parameters that influence filament size and methods to characterize it. We imaged ink filament remains and quantified some of their characteristics by changing printing speed, ink amount, and fountain solution type. Printed samples were prepared using a laboratory printability tester with varying ink levels and operating settings. Rhodamine B dye was incorporated into fountain solutions to aid in the detection of the filaments. The prints were then imaged with a confocal laser scanning microscope (CLSM) and images were further analyzed for their surface topography. Modeling of the pressure pulses in the printing nip was included to better understand the mechanism of filament formation and the origin of filament length scale. Printing speed and ink amount changed the size distribution of the observed filament remains. There was no significant difference between fountain solutions with or without isopropyl alcohol on the observed patterns of the filament remains.

ACTIVATED SLUDGE FLOC CHARACTERIZATION BY CONFOCAL LASER SCANNING MICROSCOPY

2012 ◽  
Vol 11 (3) ◽  
pp. 669-674 ◽  
Author(s):  
Szabolcs Szilveszter ◽  
Botond Raduly ◽  
Szilard Bucs ◽  
Beata Abraham ◽  
Szabolcs Lanyi ◽  
...  
Keyword(s):  
Activated Sludge ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser
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