Motility and Chemotaxis in Agrobacterium tumefaciens Surface Attachment and Biofilm Formation

Department of Biology, Indiana University, 1001 E. 3rd St., Jordan Hall 142, Bloomington, IN 47405-1847, USA.
Journal of bacteriology (Impact Factor: 2.81). 12/2007; 189(22):8005-14. DOI: 10.1128/JB.00566-07
Source: PubMed


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.

18 Reads
  • Source
    • "May 2014 | Volume 5 | Article 176 | 3 biofilm formation (Merritt et al., 2007). Ectopic expression of a plasmid-borne wild-type cheA allele enhanced motility in swim agar but did not correct the attachment deficiency. "
    [Show abstract] [Hide abstract]
    ABSTRACT: For many pathogenic bacteria surface attachment is a required first step during host interactions. Attachment can proceed to invasion of host tissue or cells or to establishment of a multicellular bacterial community known as a biofilm. The transition from a unicellular, often motile, state to a sessile, multicellular, biofilm-associated state is one of the most important developmental decisions for bacteria. Agrobacterium tumefaciens genetically transforms plant cells by transfer and integration of a segment of plasmid-encoded transferred DNA (T-DNA) into the host genome, and has also been a valuable tool for plant geneticists. A. tumefaciens attaches to and forms a complex biofilm on a variety of biotic and abiotic substrates in vitro. Although rarely studied in situ, it is hypothesized that the biofilm state plays an important functional role in the ecology of this organism. Surface attachment, motility, and cell division are coordinated through a complex regulatory network that imparts an unexpected asymmetry to the A. tumefaciens life cycle. In this review, we describe the mechanisms by which A. tumefaciens associates with surfaces, and regulation of this process. We focus on the transition between flagellar-based motility and surface attachment, and on the composition, production, and secretion of multiple extracellular components that contribute to the biofilm matrix. Biofilm formation by A. tumefaciens is linked with virulence both mechanistically and through shared regulatory molecules. We detail our current understanding of these and other regulatory schemes, as well as the internal and external (environmental) cues mediating development of the biofilm state, including the second messenger cyclic-di-GMP, nutrient levels, and the role of the plant host in influencing attachment and biofilm formation. A. tumefaciens is an important model system contributing to our understanding of developmental transitions, bacterial cell biology, and biofilm formation.
    Frontiers in Plant Science 05/2014; 5:176. DOI:10.3389/fpls.2014.00176 · 3.95 Impact Factor
  • Source
    • "The impact of chemotaxis in biofilm development depends on the bacterial species. Some works show a direct relationship between chemotaxis, attachment and biofilm development in Agrobacterium tumefaciens (Merritt et al., 2007) and Pseudomonas aeruginosa (Schmidt et al., 2011). However, in E. coli, chemotaxis was described as a non-critical process for a normal biofilm formation (Pratt & Kolter, 1998). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Xanthomonas citri spp. citri (Xcc)develops a biofilm structure both in vitro and in vivo. Despite all the progress achieved by studies regarding biofilm formation, many of its mechanisms remain poorly understood. This work focuses on the identification of new genes involved in biofilm formation and how they are related to motility, virulence and chemotaxis in Xcc. A Tn5 library of approximately 6,000 Xcc (strain 306) mutants was generated and screened to search for biofilm formation defective strains. We identified 23 genes whose association with the biofilm formation resulted in a novelty. The analysis of the 23 mutants revealed not only the involvement of new genes in biofilm formation but also reinforced the importance of exopolysaccharide production, motility and cell surface structures in this process. This collection of biofilm defective mutants underscores the multifactorial genetic program underlying the establishment of biofilm in Xcc.
    Microbiology 06/2013; 159(Pt_9). DOI:10.1099/mic.0.064709-0 · 2.56 Impact Factor
  • Source
    • "Biofilm formation can influence the interaction of A. tumefaciens with plant tissues and is tightly integrated with its physiology and morphological development. In static biofilm assays flagellar motility is required for efficient biofilm formation in A. tumefaciens [48]. The amount of adherent biomass that forms on plastic coverslips after 48 hours in static culture was measured for the kinase mutants. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The α-Proteobacterium Agrobacterium tumefaciens has proteins homologous to known regulators that govern cell division and development in Caulobacter crescentus, many of which are also conserved among diverse α-Proteobacteria. In light of recent work demonstrating similarity between the division cycle of C. crescentus and that of A. tumefaciens, the functional conservation for this presumptive control pathway was examined. In C. crescentus the CtrA response regulator serves as the master regulator of cell cycle progression and cell division. CtrA activity is controlled by an integrated pair of multi-component phosphorelays: PleC/DivJ-DivK and CckA-ChpT-CtrA. Although several of the conserved orthologues appear to be essential in A. tumefaciens, deletions in pleC or divK were isolated and resulted in cell division defects, diminished swimming motility, and a decrease in biofilm formation. A. tumefaciens also has two additional pleC/divJ homologue sensor kinases called pdhS1 and pdhS2, absent in C. crescentus. Deletion of pdhS1 phenocopied the ΔpleC and ΔdivK mutants. Cells lacking pdhS2 morphologically resembled wild-type bacteria, but were decreased in swimming motility and elevated for biofilm formation, suggesting that pdhS2 may serve to regulate the motile to non-motile switch in A. tumefaciens. Genetic analysis suggests that the PleC/DivJ-DivK and CckA-ChpT-CtrA phosphorelays in A. tumefaciens are vertically-integrated, as in C. crescentus. A gain-of-function mutation in CckA (Y674D) was identified as a spontaneous suppressor of the ΔpleC motility phenotype. Thus, although the core architecture of the A. tumefaciens pathway resembles that of C. crescentus there are specific differences including additional regulators, divergent pathway architecture, and distinct target functions.
    PLoS ONE 02/2013; 8(2):e56682. DOI:10.1371/journal.pone.0056682 · 3.23 Impact Factor
Show more

Preview (2 Sources)

18 Reads
Available from