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Spatiotemporal distribution of different extracellular polymeric substances and filamentation mediate Xylella fastidiosa adhesion and biofilm formation.

Janissen R, Murillo DM, Niza B, Sahoo PK, Nobrega MM, Cesar CL, Temperini ML, Carvalho HF, de Souza AA, Cotta MA - Sci Rep (2015)

Bottom Line: Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle.Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework.This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle.

View Article: PubMed Central - PubMed

Affiliation: Applied Physics Department, Institute of Physics 'Gleb Wataghin', State University of Campinas, 13083-859, Campinas, São Paulo, Brazil.

ABSTRACT
Microorganism pathogenicity strongly relies on the generation of multicellular assemblies, called biofilms. Understanding their organization can unveil vulnerabilities leading to potential treatments; spatially and temporally-resolved comprehensive experimental characterization can provide new details of biofilm formation, and possibly new targets for disease control. Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle. Single cell adhesion is caused by unspecific electrostatic interactions through proteins at the cell polar region, where EPS accumulation is required for more firmly-attached, irreversibly adhered cells. Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework. During biofilm maturation, soluble EPS forms a filamentous matrix that facilitates cell adhesion and provides mechanical support, while the biofilm keeps anchored by few cells. This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle. Using new approaches, our findings provide insights regarding different aspects of the adhesion process of X. fastidiosa and biofilm formation.

No MeSH data available.


Related in: MedlinePlus

Biofilm architecture and growing network of S-EPS film and filamentous EPS structures.(a) Ex-vivo SDCLM data of a mid-sized, vertical growing biofilm. The bottom view of this small biofilm indicates a relatively small number of surface-adhered cells (light green color). (b) False-color WFM and SDCLM fluorescence images identify S-EPS film surrounding a bacterial biofilm and filamentous EPS structures. (c) False-color WFM fluorescence data showing filamentous structures emerging from bacterial biofilm and facilitated bacterial adhesion on the filamentous network. (d) SPM and ex-vivo SDLCM data illustrate bacterial adhesion to filamentous EPS structures. (e) AFM topography data of filamentous EPS structures and complementary height distribution of individual EPS filaments (n = 89). See also Supplementary Fig. S4. Measurement statistics are described in the Materials and Methods section.
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f4: Biofilm architecture and growing network of S-EPS film and filamentous EPS structures.(a) Ex-vivo SDCLM data of a mid-sized, vertical growing biofilm. The bottom view of this small biofilm indicates a relatively small number of surface-adhered cells (light green color). (b) False-color WFM and SDCLM fluorescence images identify S-EPS film surrounding a bacterial biofilm and filamentous EPS structures. (c) False-color WFM fluorescence data showing filamentous structures emerging from bacterial biofilm and facilitated bacterial adhesion on the filamentous network. (d) SPM and ex-vivo SDLCM data illustrate bacterial adhesion to filamentous EPS structures. (e) AFM topography data of filamentous EPS structures and complementary height distribution of individual EPS filaments (n = 89). See also Supplementary Fig. S4. Measurement statistics are described in the Materials and Methods section.

Mentions: A thorough scrutiny of the biofilm in maturation (depicted in ex-vivo SDCLM images, Fig. 4a) illuminates several aspects of its architecture. After ~72 h of cell growth, mature biofilms continue to exhibit enhanced growth in the vertical direction (Fig. 4a) with thicknesses >40 µm; however, no predominant areas for vertical growth are furthermore observable. Similarly, as observed previously for young biofilms, the entire biofilm structure is anchored to the surface only by a relatively small number of cells, reminiscent of the original irreversibly attached cells (Fig. 4a, bottom view and Supplementary Movie S4).


Spatiotemporal distribution of different extracellular polymeric substances and filamentation mediate Xylella fastidiosa adhesion and biofilm formation.

Janissen R, Murillo DM, Niza B, Sahoo PK, Nobrega MM, Cesar CL, Temperini ML, Carvalho HF, de Souza AA, Cotta MA - Sci Rep (2015)

Biofilm architecture and growing network of S-EPS film and filamentous EPS structures.(a) Ex-vivo SDCLM data of a mid-sized, vertical growing biofilm. The bottom view of this small biofilm indicates a relatively small number of surface-adhered cells (light green color). (b) False-color WFM and SDCLM fluorescence images identify S-EPS film surrounding a bacterial biofilm and filamentous EPS structures. (c) False-color WFM fluorescence data showing filamentous structures emerging from bacterial biofilm and facilitated bacterial adhesion on the filamentous network. (d) SPM and ex-vivo SDLCM data illustrate bacterial adhesion to filamentous EPS structures. (e) AFM topography data of filamentous EPS structures and complementary height distribution of individual EPS filaments (n = 89). See also Supplementary Fig. S4. Measurement statistics are described in the Materials and Methods section.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4402645&req=5

f4: Biofilm architecture and growing network of S-EPS film and filamentous EPS structures.(a) Ex-vivo SDCLM data of a mid-sized, vertical growing biofilm. The bottom view of this small biofilm indicates a relatively small number of surface-adhered cells (light green color). (b) False-color WFM and SDCLM fluorescence images identify S-EPS film surrounding a bacterial biofilm and filamentous EPS structures. (c) False-color WFM fluorescence data showing filamentous structures emerging from bacterial biofilm and facilitated bacterial adhesion on the filamentous network. (d) SPM and ex-vivo SDLCM data illustrate bacterial adhesion to filamentous EPS structures. (e) AFM topography data of filamentous EPS structures and complementary height distribution of individual EPS filaments (n = 89). See also Supplementary Fig. S4. Measurement statistics are described in the Materials and Methods section.
Mentions: A thorough scrutiny of the biofilm in maturation (depicted in ex-vivo SDCLM images, Fig. 4a) illuminates several aspects of its architecture. After ~72 h of cell growth, mature biofilms continue to exhibit enhanced growth in the vertical direction (Fig. 4a) with thicknesses >40 µm; however, no predominant areas for vertical growth are furthermore observable. Similarly, as observed previously for young biofilms, the entire biofilm structure is anchored to the surface only by a relatively small number of cells, reminiscent of the original irreversibly attached cells (Fig. 4a, bottom view and Supplementary Movie S4).

Bottom Line: Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle.Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework.This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle.

View Article: PubMed Central - PubMed

Affiliation: Applied Physics Department, Institute of Physics 'Gleb Wataghin', State University of Campinas, 13083-859, Campinas, São Paulo, Brazil.

ABSTRACT
Microorganism pathogenicity strongly relies on the generation of multicellular assemblies, called biofilms. Understanding their organization can unveil vulnerabilities leading to potential treatments; spatially and temporally-resolved comprehensive experimental characterization can provide new details of biofilm formation, and possibly new targets for disease control. Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle. Single cell adhesion is caused by unspecific electrostatic interactions through proteins at the cell polar region, where EPS accumulation is required for more firmly-attached, irreversibly adhered cells. Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework. During biofilm maturation, soluble EPS forms a filamentous matrix that facilitates cell adhesion and provides mechanical support, while the biofilm keeps anchored by few cells. This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle. Using new approaches, our findings provide insights regarding different aspects of the adhesion process of X. fastidiosa and biofilm formation.

No MeSH data available.


Related in: MedlinePlus