scholarly journals Motility and Chemotaxis in Agrobacterium tumefaciens Surface Attachment and Biofilm Formation

2007 ◽  
Vol 189 (22) ◽  
pp. 8005-8014 ◽  
Author(s):  
Peter M. Merritt ◽  
Thomas Danhorn ◽  
Clay Fuqua
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Biofilm Formation ◽  
Bacterial Species ◽  
Wild Type ◽  
Swarming Motility ◽  
Surface Attachment ◽  
Directed Motility ◽  
Suppressor Mutants ◽  
Static Conditions ◽  
Specific Deletion

ABSTRACT Bacterial motility mechanisms, including swimming, swarming, and twitching, are known to have important roles in biofilm formation, including colonization and the subsequent expansion into mature structured surface communities. Directed motility requires chemotaxis functions that are conserved among many bacterial species. The biofilm-forming plant pathogen Agrobacterium tumefaciens drives swimming motility by utilizing a small group of flagella localized to a single pole or the subpolar region of the cell. There is no evidence for twitching or swarming motility in A. tumefaciens. Site-specific deletion mutations that resulted in either aflagellate, flagellated but nonmotile, or flagellated but nonchemotactic A. tumefaciens derivatives were examined for biofilm formation under static and flowing conditions. Nonmotile mutants were significantly deficient in biofilm formation under static conditions. Under flowing conditions, however, the aflagellate mutant rapidly formed aberrantly dense, tall biofilms. In contrast, a nonmotile mutant with unpowered flagella was clearly debilitated for biofilm formation relative to the wild type. A nontumbling chemotaxis mutant was only weakly affected with regard to biofilm formation under nonflowing conditions but was notably compromised in flow, generating less adherent biomass than the wild type, with a more dispersed cellular arrangement. Extragenic suppressor mutants of the chemotaxis-impaired, straight-swimming phenotype were readily isolated from motility agar plates. These mutants regained tumbling at a frequency similar to that of the wild type. Despite this phenotype, biofilm formation by the suppressor mutants in static cultures was significantly deficient. Under flowing conditions, a representative suppressor mutant manifested a phenotype similar to yet distinct from that of its nonchemotactic parent.

scholarly journals Advanced understanding of prokaryotic biofilm formation using a cost-effective and versatile multi-panel adhesion (mPAD) mount

2022 ◽  
Author(s):  
Stefan Schulze ◽  
Heather Schiller ◽  
Jordan Solomonic ◽  
Orkan Telhan ◽  
Kyle Costa ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Cost Effective ◽  
Flow Chamber ◽  
Culture Conditions ◽  
Multiple Time ◽  
Wild Type ◽  
Surface Attachment ◽  
Flowing Liquid ◽  
Multiple Time Points ◽  
Static Conditions

Most microorganisms exist in biofilms, which comprise aggregates of cells surrounded by an extracellular matrix that provides protection from external stresses. Based on the conditions under which they form, biofilm structures vary in significant ways. For instance, biofilms that develop when microbes are incubated under static conditions differ from those formed when microbes encounter the shear forces of a flowing liquid. Moreover, biofilms develop dynamically over time. Here, we describe a cost-effective, 3D-printed coverslip holder that facilitates surface adhesion assays under a broad range of standing and shaking culture conditions. This multi-panel adhesion (mPAD) mount further allows cultures to be sampled at multiple time points, ensuring consistency and comparability between samples and enabling analyses of the dynamics of biofilm formation. As a proof of principle, using the mPAD mount for shaking, oxic cultures, we confirm previous flow chamber experiments showing that Pseudomonas aeruginosa wild type and a phenazine deletion mutant (Δ phz ) form biofilms with similar structure but reduced density in the mutant strain. Extending this analysis to anoxic conditions, we reveal that microcolony and biofilm formation can only be observed under shaking conditions and are decreased in the Δ phz mutant compared to wild-type cultures, indicating that phenazines are crucial for the formation of biofilms if oxygen as an electron acceptor is unavailable. Furthermore, while the model archaeon Haloferax volcanii does not require archaella for surface attachment under static conditions, we demonstrate that H. volcanii mutants that lack archaella are impaired in early stages of biofilm formation under shaking conditions. Importance: Due to the versatility of the mPAD mount, we anticipate that it will aid the analysis of biofilm formation in a broad range of bacteria and archaea. Thereby, it contributes to answering critical biological questions about the regulatory and structural components of biofilm formation and understanding this process in a wide array of environmental, biotechnological, and medical contexts.

scholarly journals Truncation in the core oligosaccharide of lipopolysaccharide affects flagella-mediated motility in Pseudomonas aeruginosa PAO1 via modulation of cell surface attachment

Microbiology ◽  
2009 ◽  
Vol 155 (10) ◽  
pp. 3449-3460 ◽  
Author(s):  
Theresa Lindhout ◽  
Peter C. Y. Lau ◽  
Dyanne Brewer ◽  
Joseph S. Lam
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Surface ◽  
Bacterial Species ◽  
Soft Agar ◽  
Wild Type ◽  
Core Oligosaccharide ◽  
Swarming Motility ◽  
Surface Attachment ◽  
Abiotic Surfaces ◽  
Isogenic Mutants

In many Gram-negative bacterial species, rough strains producing truncated lipopolysaccharide (LPS) generally exhibit defects in motility compared with smooth strains. However, the role that LPS plays in bacterial motility is not well understood. The goal of this study was to examine the relationship between LPS defects and motility of Pseudomonas aeruginosa. P. aeruginosa wild-type strain PAO1 and three isogenic mutants with defects in the rmlC, migA and wapR genes and producing truncated core oligosaccharide were investigated in terms of motility, attachment to glass and flagella expression. Compared with the wild-type, the three mutants showed significant retardation in both swarming motility on 0.5 % soft-agar plates and swimming motility on 0.3 % soft-agar plates. Moreover, attachment to abiotic surfaces was observed to be stronger in these mutants. The assembly of flagella appeared to be intact in these strains and the ability of individual cells to swim was unaffected. Flagellin proteins prepared from mutants rmlC and rmd, defective in the production of TDP-l-rhamnose and GDP-d-rhamnose, respectively, were compared and a change in molecular mass was observed only in the rmlC mutant. These data indicated that l-rhamnose, and not its enantiomer, d-rhamnose, is incorporated into the flagellin glycan of P. aeruginosa PAO1. The nucleotide-activated sugar precursor TDP-l-rhamnose is therefore shared between LPS biosynthesis and flagellin glycosylation in P. aeruginosa PAO1. Our results suggest that although biochemical precursors are shared by LPS and flagellin glycan biosynthesis, LPS truncations probably alter flagella-mediated motility in P. aeruginosa by modulating cell-surface attachment but not flagella synthesis.

scholarly journals Effect of Nitroxides on Swarming Motility and Biofilm Formation, Multicellular Behaviors in Pseudomonas aeruginosa

2013 ◽  
Vol 57 (10) ◽  
pp. 4877-4881 ◽  
Author(s):  
César de la Fuente-Núñez ◽  
Fany Reffuveille ◽  
Kathryn E. Fairfull-Smith ◽  
Robert E. W. Hancock
Keyword(s):  
Nitric Oxide ◽  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Wild Type ◽  
Planktonic Cells ◽  
Swarming Motility ◽  
Content Type ◽  
Exogenous Addition ◽  
Biofilm Dispersion ◽  
Dispersal Activity

ABSTRACTThe ability of nitric oxide (NO) to induce biofilm dispersion has been well established. Here, we investigated the effect of nitroxides (sterically hindered nitric oxide analogues) on biofilm formation and swarming motility inPseudomonas aeruginosa. A transposon mutant unable to produce nitric oxide endogenously (nirS) was deficient in swarming motility relative to the wild type and the complemented strain. Moreover, expression of thenirSgene was upregulated by 9.65-fold in wild-type swarming cells compared to planktonic cells. Wild-type swarming levels were substantially restored upon the exogenous addition of nitroxide containing compounds, a finding consistent with the hypothesis that NO is necessary for swarming motility. Here, we showed that nitroxides not only mimicked the dispersal activity of NO but also prevented biofilms from forming in flow cell chambers. In addition, anirStransposon mutant was deficient in biofilm formation relative to the wild type and the complemented strain, thus implicating NO in the formation of biofilms. Intriguingly, despite its stand-alone action in inhibiting biofilm formation and promoting dispersal, a nitroxide partially restored the ability of anirSmutant to form biofilms.

scholarly journals Spermidine Inversely Influences Surface Interactions and Planktonic Growth in Agrobacterium tumefaciens

2016 ◽  
Vol 198 (19) ◽  
pp. 2682-2691 ◽  
Author(s):  
Yi Wang ◽  
Sok Ho Kim ◽  
Ramya Natarajan ◽  
Jason E. Heindl ◽  
Eric L. Bruger ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Biofilm Formation ◽  
Direct Synthesis ◽  
Wild Type ◽  
Cyclic Diguanylate ◽  
Content Type ◽  
Exogenous Addition ◽  
In The Wild ◽  
Growth Requirement ◽  
Transposon Insertions

ABSTRACTIn bacteria, the functions of polyamines, small linear polycations, are poorly defined, but these metabolites can influence biofilm formation in several systems. Transposon insertions in an ornithine decarboxylase (odc) gene inAgrobacterium tumefaciens, predicted to direct synthesis of the polyamine putrescine from ornithine, resulted in elevated cellulose. Null mutants forodcgrew somewhat slowly in a polyamine-free medium but exhibited increased biofilm formation that was dependent on cellulose production. Spermidine is an essential metabolite inA. tumefaciensand is synthesized from putrescine inA. tumefaciensvia the stepwise actions of carboxyspermidine dehydrogenase (CASDH) and carboxyspermidine decarboxylase (CASDC). Exogenous addition of either putrescine or spermidine to theodcmutant returned biofilm formation to wild-type levels. Low levels of exogenous spermidine restored growth to CASDH and CASDC mutants, facilitating weak biofilm formation, but this was dampened with increasing concentrations. Norspermidine rescued growth for theodc, CASDH, and CASDC mutants but did not significantly affect their biofilm phenotypes, whereas in the wild type, it stimulated biofilm formation and depressed spermidine levels. Theodcmutant produced elevated levels of cyclic diguanylate monophosphate (c-di-GMP), exogenous polyamines modulated these levels, and expression of a c-di-GMP phosphodiesterase reversed the enhanced biofilm formation. Prior work revealed accumulation of the precursors putrescine and carboxyspermidine in the CASDH and CASDC mutants, respectively, but unexpectedly, both mutants accumulated homospermidine; here, we show that this requires a homospermidine synthase (hss) homologue.IMPORTANCEPolyamines are small, positively charged metabolites that are nearly ubiquitous in cellular life. They are often essential in eukaryotes and more variably in bacteria. Polyamines have been reported to influence the surface-attached biofilm formation of several bacteria. InAgrobacterium tumefaciens, mutants with diminished levels of the polyamine spermidine are stimulated for biofilm formation, and exogenous provision of spermidine decreases biofilm formation. Spermidine is also essential forA. tumefaciensgrowth, but the related polyamine norspermidine exogenously rescues growth and does not diminish biofilm formation, revealing that the growth requirement and biofilm control are separable. Polyamine control of biofilm formation appears to function via effects on the cellular second messenger cyclic diguanylate monophosphate, regulating the transition from a free-living to a surface-attached lifestyle.

scholarly journals Quorum Sensing Influences Burkholderia thailandensis Biofilm Development and Matrix Production

2016 ◽  
Vol 198 (19) ◽  
pp. 2643-2650 ◽  
Author(s):  
Boo Shan Tseng ◽  
Charlotte D. Majerczyk ◽  
Daniel Passos da Silva ◽  
Josephine R. Chandler ◽  
E. Peter Greenberg ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Biofilm Formation ◽  
Bacterial Species ◽  
Matrix Material ◽  
Close Relative ◽  
Wild Type ◽  
Flow Conditions ◽  
Burkholderia Thailandensis ◽  
Content Type ◽  
Biofilm Development

ABSTRACTMembers of the genusBurkholderiaare known to be adept at biofilm formation, which presumably assists in the survival of these organisms in the environment and the host. Biofilm formation has been linked to quorum sensing (QS) in several bacterial species. In this study, we characterizedBurkholderia thailandensisbiofilm development under flow conditions and sought to determine whether QS contributes to this process.B. thailandensisbiofilm formation exhibited an unusual pattern: the cells formed small aggregates and then proceeded to produce mature biofilms characterized by “dome” structures filled with biofilm matrix material. We showed that this process was dependent on QS.B. thailandensishas three acyl-homoserine lactone (AHL) QS systems (QS-1, QS-2, and QS-3). An AHL-negative strain produced biofilms consisting of cell aggregates but lacking the matrix-filled dome structures. This phenotype was rescued via exogenous addition of the three AHL signals. Of the threeB. thailandensisQS systems, we show that QS-1 is required for proper biofilm development, since abtaR1mutant, which is defective in QS-1 regulation, forms biofilms without these dome structures. Furthermore, our data show that the wild-type biofilm biomass, as well as the material inside the domes, stains with a fucose-binding lectin. ThebtaR1mutant biofilms, however, are negative for fucose staining. This suggests that the QS-1 system regulates the production of a fucose-containing exopolysaccharide in wild-type biofilms. Finally, we present data showing that QS ability during biofilm development produces a biofilm that is resistant to dispersion under stress conditions.IMPORTANCEThe saprophyteBurkholderia thailandensisis a close relative of the pathogenic bacteriumBurkholderia pseudomallei, the causative agent of melioidosis, which is contracted from its environmental reservoir. Since most bacteria in the environment reside in biofilms,B. thailandensisis an ideal model organism for investigating questions inBurkholderiaphysiology. In this study, we characterizedB. thailandensisbiofilm development and sought to determine if quorum sensing (QS) contributes to this process. Our work shows thatB. thailandensisproduces biofilms with unusual dome structures under flow conditions. Our findings suggest that these dome structures are filled with a QS-regulated, fucose-containing exopolysaccharide that may be involved in the resilience ofB. thailandensisbiofilms against changes in the nutritional environment.

scholarly journals Exopolysaccharide Production Is Required for Biofilm Formation and Plant Colonization by the Nitrogen-Fixing Endophyte Gluconacetobacter diazotrophicus

2011 ◽  
Vol 24 (12) ◽  
pp. 1448-1458 ◽  
Author(s):  
Carlos H. S. G. Meneses ◽  
Luc F. M. Rouws ◽  
Jean L. Simões-Araújo ◽  
Marcia S. Vidal ◽  
José I. Baldani
Keyword(s):  
Biofilm Formation ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Normal Growth ◽  
Root Surface ◽  
Plant Colonization ◽  
Gluconacetobacter Diazotrophicus ◽  
Nitrogen Fixing ◽  
Wild Type ◽  
Endophytic Colonization

The genome of the endophytic diazotrophic bacterial species Gluconacetobacter diazotrophicus PAL5 (PAL5) revealed the presence of a gum gene cluster. In this study, the gumD gene homologue, which is predicted to be responsible for the first step in exopolysaccharide (EPS) production, was insertionally inactivated and the resultant mutant (MGD) was functionally studied. The mutant MGD presented normal growth and nitrogen (N2) fixation levels but did not produce EPS when grown on different carbon sources. MGD presented altered colony morphology on soft agar plates (0.3% agar) and was defective in biofilm formation on glass wool. Most interestingly, MGD was defective in rice root surface attachment and in root surface and endophytic colonization. Genetic complementation reverted all mutant phenotypes. Also, the addition of EPS purified from culture supernatants of the wild-type strain PAL5 to the mutant MGD was effective in partially restoring wild-type biofilm formation and plant colonization. These data provide strong evidence that the PAL5 gumD gene is involved in EPS biosynthesis and that EPS biosynthesis is required for biofilm formation and plant colonization. To our knowledge, this is the first report of a role of EPS in the endophytic colonization of graminaceous plants by a nitrogen-fixing bacterium.

scholarly journals IscR Controls Iron-Dependent Biofilm Formation in Escherichia coli by Regulating Type I Fimbria Expression

2008 ◽  
Vol 191 (4) ◽  
pp. 1248-1257 ◽  
Author(s):  
Yun Wu ◽  
F. Wayne Outten
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Developmental Process ◽  
Bacterial Species ◽  
Type I ◽  
Surface Attachment ◽  
Environmental Signals ◽  
Regulatory Pathways ◽  
E Coli ◽  
Type I Fimbriae

ABSTRACT Biofilm formation is a complex developmental process regulated by multiple environmental signals. In addition to other nutrients, the transition metal iron can also regulate biofilm formation. Iron-dependent regulation of biofilm formation varies by bacterial species, and the exact regulatory pathways that control iron-dependent biofilm formation are often unknown or only partially characterized. To address this gap in our knowledge, we examined the role of iron availability in regulating biofilm formation in Escherichia coli. The results indicate that biofilm formation is repressed under low-iron conditions in E. coli. Furthermore, a key iron regulator, IscR, controls biofilm formation in response to changes in cellular Fe-S homeostasis. IscR regulates the FimE recombinase to control expression of type I fimbriae in E. coli. We propose that iron-dependent regulation of FimE via IscR leads to decreased surface attachment and biofilm dispersal under iron-limiting conditions.

EFFECT OF TOPOGRAPHICALLY PATTERNED POLY(DIMETHYLSILOXANE) SURFACES ON Pseudomonas aeruginosa ADHESION AND BIOFILM FORMATION

Nano LIFE ◽  
2012 ◽  
Vol 02 (04) ◽  
pp. 1242004 ◽  
Author(s):  
JOHN F. LING ◽  
MARY V. GRAHAM ◽  
NATHANIEL C. CADY
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Lower Surface ◽  
Bacterial Attachment ◽  
Initial Surface ◽  
Direct Role ◽  
Surface Attachment ◽  
Bacterial Surface ◽  
Static Conditions

Bacterial pathogens, such as Pseudomonas aeruginosa, readily form biofilms on surfaces, limiting the efficacy of antimicrobial and antibiotic treatments. To mitigate biofilm formation, surfaces are often treated with antimicrobial agents, which have limited lifetime and efficacy. Recent studies have shown that well-ordered topographic patterns can limit bacterial attachment to surfaces and limit biofilm formation. In this study, nano and microscale patterned poly(dimethylsiloxane) surfaces were evaluated for their ability to affect adhesion and biofilm formation by Pseudomonas aeruginosa. Feature size and spacing were varied from 500 nm to 2 μm and included repeating arrays of square pillars, holes, lines and biomimetc Sharklet™ patterns. Bacterial surface adhesion and biofilm formation was assessed in microfluidic flow devices and under static conditions. Attachment profiles under static and fluid flow varied within topography types, sizes and spacing. Pillar structures of all sizes yielded lower surface attachment than line-based patterns and arrays of holes. This trend was also observed for biomimetic Sharklet™ patterns, with reduced bacterial attachment to "raised" features as compared to "recessed" features. Notably, none of the topographically patterned surfaces outperformed smooth surfaces (without topography) for resisting cell adhesion. Initial surface attachment patterns were indicative of subsequent biofilm formation and coverage, suggesting a direct role of surface topography in biofilm-based biofouling.

scholarly journals A Rhizobiales-Specific Unipolar Polysaccharide Adhesin Contributes to Rhodopseudomonas palustris Biofilm Formation across Diverse Photoheterotrophic Conditions

2016 ◽  
Vol 83 (4) ◽  
Author(s):  
Ryan K. Fritts ◽  
Breah LaSarre ◽  
Ari M. Stoner ◽  
Amanda L. Posto ◽  
James B. McKinlay
Keyword(s):  
Biofilm Formation ◽  
Bacterial Species ◽  
Rhodopseudomonas Palustris ◽  
Gene Clusters ◽  
Growth Conditions ◽  
Organic Substrates ◽  
Biosynthesis Gene Cluster ◽  
Surface Attachment ◽  
Content Type ◽  
Animal Pathogens

ABSTRACT Bacteria predominantly exist as members of surfaced-attached communities known as biofilms. Many bacterial species initiate biofilms and adhere to each other using cell surface adhesins. This is the case for numerous ecologically diverse Alphaprotebacteria, which use polar exopolysaccharide adhesins for cell-cell adhesion and surface attachment. Here, we show that Rhodopseudomonas palustris, a metabolically versatile member of the alphaproteobacterial order Rhizobiales, contains a functional unipolar polysaccharide (UPP) biosynthesis gene cluster. Deletion of genes predicted to be critical for UPP biosynthesis and export abolished UPP production. We also found that R. palustris uses UPP to mediate biofilm formation across diverse photoheterotrophic growth conditions, wherein light and organic substrates are used to support growth. However, UPP was less important for biofilm formation during photoautotrophy, where light and CO2 support growth, and during aerobic respiration with organic compounds. Expanding our analysis beyond R. palustris, we examined the phylogenetic distribution and genomic organization of UPP gene clusters among Rhizobiales species that inhabit diverse niches. Our analysis suggests that UPP is a conserved ancestral trait of the Rhizobiales but that it has been independently lost multiple times during the evolution of this clade, twice coinciding with adaptation to intracellular lifestyles within animal hosts. IMPORTANCE Bacteria are ubiquitously found as surface-attached communities and cellular aggregates in nature. Here, we address how bacterial adhesion is coordinated in response to diverse environments using two complementary approaches. First, we examined how Rhodopseudomonas palustris, one of the most metabolically versatile organisms ever described, varies its adhesion to surfaces in response to different environmental conditions. We identified critical genes for the production of a unipolar polysaccharide (UPP) and showed that UPP is important for adhesion when light and organic substrates are used for growth. Looking beyond R. palustris, we performed the most comprehensive survey to date on the conservation of UPP biosynthesis genes among a group of closely related bacteria that occupy diverse niches. Our findings suggest that UPP is important for free-living and plant-associated lifestyles but dispensable for animal pathogens. Additionally, we propose guidelines for classifying the adhesins produced by various Alphaprotebacteria, facilitating future functional and comparative studies.

Alternative Sigma Factor σB Is Not Essential for Listeria monocytogenes Surface Attachment

2005 ◽  
Vol 68 (2) ◽  
pp. 311-317 ◽  
Author(s):  
UTE SCHWAB ◽  
YUEWEI HU ◽  
MARTIN WIEDMANN ◽  
KATHRYN J. BOOR
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Single Cells ◽  
Foodborne Pathogen ◽  
Plate Count ◽  
Initial Surface ◽  
Wild Type ◽  
Surface Attachment ◽  
Steel Analysis ◽  
Static Conditions

Listeria monocytogenes is a foodborne pathogen frequently isolated from the food processing environment. Multiple lines of evidence suggested a possible role for the L. monocytogenes alternative transcription factor sigma B (σB) in surface attachment and biofilm formation. Therefore, through plate count and microscopic techniques, the L. monocytogenes 10403S strain and an otherwise isogenic ΔsigB strain were tested for attachment to stainless steel. Analysis of microscopic images revealed that after 72 h at 24°C under static conditions the tested L. monocytogenes strains attached uniformly to surfaces as single cells. Both strains were capable of rapid attachment (i.e., numbers of attached cells were essentially the same after either 5 min or 24 h of incubation). Numbers of attached ΔsigB cells were significantly lower than those of the wild-type strain after 48 and 72 h of incubation at 24°C (P = 0.001). Similar numbers of the ΔsigB strain attached to stainless steel regardless of temperature (24 or 37°C); however, ΔsigB cells attached at higher relative numbers in the presence of 6% NaCl after 48 and 72 h. Furthermore, in the presence of Pseudomonas fluorescens, similarly high numbers of wild-type and ΔsigB cells attached to the surfaces, forming mixed biofilms. Our data suggest that σB is not required for initial surface attachment of L. monocytogenes.

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