Bacterial Growth Cycle Research Articles

Computational and in vitro analyses of the antibacterial effect of the ethanolic extract of Pluchea indica L. leaves.

The most common gram-negative, Escherichia coli, and gram-positive bacteria, Bacillus spp., have evolved different mechanisms that have caused the emergence of multi-drug resistance. As a result, drugs that block the bacterial growth cycle are needed. Here, in silico and in vitro studies were performed to assess compounds in the Pluchea indica leaf extract, a medicinal plant, that can inhibit bacterial proteins. Briefly, P. indica leaves were extracted using ethanol. The crude extract was then subjected to gas chromatography-mass spectrometry for metabolite screening. Molecular docking simulations with rhomboid protease (Rpro) (Protein data bank ID number: 3ZMI from E. coli and filamenting temperature-sensitive mutant Z (FtsZ) protein data bank ID number: 2VAM from Bacillus subtilis were performed. Moreover, the well diffusion method was used to confirm the antibacterial activity of P. indica leaf extract. A total of 10 compounds were identified in the P. indica extract and used for computational analysis. Based on drug-likeness prediction, P. indica compounds may be drug-like molecules. Binding affinity tests indicated that 10,10-Dimethyl-2,6-dimethylenebicyclo(7.2.0)undecan-5.β.-ol and 11,11-Dimethyl-4,8-dimethylenebicyclo(7.2.0)undecan-3-ol had the most negative values. Accordingly, these compounds may be potential ligands that bind to bacterial proteins. The root mean square fluctuation values was <2 Å, indicating stable fluctuation binding for the ligand-protein complex. According to in vitro antibacterial assays, a high concentration (50%) of the P. indica extract markedly inhibited E. coli and B. subtilis, with inhibitory zone diameters of 31.86±1.63 and 21.09±0.09 mm, respectively. Overall, the compounds in the P. indica leaf extract were identified as functional inhibitors of E. coli and B. subtilis proteins via in silico analysis. This may facilitate development of antibacterial agents.

Spatiotemporal Analysis of Secondary Metabolites Produced by S.coelicolor Using Automated Sampling Mass Spectrometry

Natural products (NPs) are organic compounds that are synthesized by living organisms such as bacteria. NPs and their derivatives are very useful in the medical field to develop new drugs, particularly new antibiotics which are crucial to combat developing antibiotic resistance. The current research on the secondary metabolites (SMs) produced throughout the growth cycle of bacteria is limited, as the standard procedure is to examine the SM production at the end of the bacteria’s growth cycle. Therefore, there may be many undiscovered SMs which are produced and consumed prior to analysis. A mass spectrometer (MS) can be used to track analytes of interest, such as SMs. When the MS is paired with an ambient ionization technique like the Liquid Micro-Junction Surface Sampling Probe (LMJ-SSP) it works as a minimally invasive approach to track the SMs in real time produced throughout the bacterial growth cycle. This allows for the SMs to be tracked both temporally and spatially without killing the bacteria. The process is automated using the LMJ-SSP mounted to a 3D printer, allowing for a grid-like sampling process to map NPs production. Herein, A 3D-printed growth chamber dubbed the “Washing And Sampling Agar Bacterial Inoculation” (WASABI) Box was designed to resemble a sampling grid for simpler analysis. Another key feature of the WASABI Box are washing ports, containing 70% ethanol, to sterilize the LMJ-SSP and minimize contamination between rows. The SM production of S. coelicolor in the WASABI Box was analyzed from day 4 to 7 of the bacterial growth cycle, where the data was used to generate spatial-temporal heat maps tracking metabolites including Albaflavenone (m/z 219.2), Desferrioxamine B (m/z 561.4), Germicidin B (m/z 183.1), Kalafungin (m/z 301.1), and Streptorubin B (m/z 392.3).

The mitigation potential of synergistic quorum quenching and antibacterial properties for biofilm proliferation and membrane biofouling

Biofouling has been a persistent problem hindering the application of membranes in water treatment, and quorum quenching has been identified as an effective method for mitigating biofouling, but surface accumulation of live bacteria still induces biofilm secretion, which poses a significant challenge for sustained prevention of membrane biofouling. In this study, we utilized quercetin, a typical flavonoid with the dual functions of quorum quenching and bacterial inactivation, to evaluate its role in preventing biofilm proliferation and against biofouling. Quercetin exhibited excellent antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), and the decreased bioactivity was positively correlated with the quercetin concentration, with inhibition rates of 53.1 % and 57.4 %, respectively, at the experimental concentrations. The RT-qPCR results demonstrated that quercetin inhibited AI-2 of E. coli and AGR of S. aureus mediated quorum sensing system, and reduced the expression of genes such as adhesion, virulence, biofilm secretion, and key regulatory proteases. As a result, the bacterial growth cycle was retarded and the biomass and biofilm maturation cycles were alleviated with the synergistic effect of quorum quenching and antibacterial activity. In addition, membrane biofouling was significantly declined in the dynamic operation experiments, dead cells in the biofilm overwhelmingly dominated, and the final normalized water fluxes were increased by more than 49.9 % and 34.5 % for E. coli and S. aureus, respectively. This work demonstrates the potential for mitigating biofouling using protocols that quorum quenching and inactivate bacteria, also provides a unique and long-lasting strategy to alleviate membrane fouling.

N-Acylated Ciprofloxacin Derivatives: Synthesis and In Vitro Biological Evaluation as Antibacterial and Anticancer Agents.

A novel series of N-acylated ciprofloxacin (CP) conjugates 1-21 were synthesized and screened as potential antimicrobial agents. Conjugates 1 and 2 were 1.25-10-fold more potent than CP toward all Staphylococci (minimal inhibitory concentration 0.05-0.4 μg/mL). Most of the chloro- (3-7), bromo- (8-11), and CF3-alkanoyl (14-16) derivatives expressed higher or comparable activity to CP against selected Gram-positive strains. A few CP analogues (5, 10, and 11) were also more effective toward the chosen clinical Gram-negative rods. Conjugates 5, 10, and 11 considerably influenced the phases of the bacterial growth cycle over 18 h. Additionally, compounds 2, 4-7, 9-12, and 21 exerted stronger tuberculostatic action against three Mycobacterium tuberculosis isolates than the first-line antitubercular drugs. Amides 1, 2, 5, 6, 10, and 11 targeted gyrase and topoisomerase IV at 2.7-10.0 μg/mL, which suggests a mechanism of antibacterial action related to CP. These findings were confirmed by molecular docking studies. In addition, compounds 3 and 15 showed high antiproliferative activities against prostate PC3 cells (IC50 2.02-4.8 μM), up to 6.5-2.75 stronger than cisplatin. They almost completely reduced the growth and proliferation rates in these cells, without a cytotoxic action against normal HaCaT cell lines. Furthermore, derivatives 3 and 21 induced apoptosis/necrosis in PC3 cells, probably by increasing the intracellular ROS amount, as well as they diminished the IL-6 level in tumor cells.

Spatiotemporal Coupling of DNA Supercoiling and Genomic Sequence Organization—A Timing Chain for the Bacterial Growth Cycle?

In this article we describe the bacterial growth cycle as a closed, self-reproducing, or autopoietic circuit, reestablishing the physiological state of stationary cells initially inoculated in the growth medium. In batch culture, this process of self-reproduction is associated with the gradual decline in available metabolic energy and corresponding change in the physiological state of the population as a function of “travelled distance” along the autopoietic path. We argue that this directional alteration of cell physiology is both reflected in and supported by sequential gene expression along the chromosomal OriC-Ter axis. We propose that during the E. coli growth cycle, the spatiotemporal order of gene expression is established by coupling the temporal gradient of supercoiling energy to the spatial gradient of DNA thermodynamic stability along the chromosomal OriC-Ter axis.

A motile doublet form of Salmonella Typhimurium diversifies target search behavior at the epithelial surface.

The behaviors of infectious bacteria are commonly studied in bulk. This is effective to define the general properties of a given isolate, but insufficient to resolve subpopulations and unique single‐microbe behaviors within the bacterial pool. We here employ microscopy to study single‐bacterium characteristics among Salmonella enterica serovar Typhimurium (S.Tm), as they prepare for and launch invasion of epithelial host cells. We find that during the bacterial growth cycle, S.Tm populations switch gradually from fast planktonic growth to a host cell‐invasive phenotype, characterized by flagellar motility and expression of the Type‐three‐secretion‐system‐1. The indistinct nature of this shift leads to the establishment of a transient subpopulation of S.Tm “doublets”—waist‐bearing bacteria anticipating cell division—which simultaneously express host cell invasion machinery. In epithelial cell culture infections, these S.Tm doublets outperform their “singlet” brethren and represent a hyperinvasive subpopulation. Atop both glass and enteroid‐derived monolayers, doublets swim along markedly straighter trajectories than singlets, thereby diversifying search patterns and improving the surface exploration capacity of the total bacterial population. The straighter swimming, combined with an enhanced cell‐adhesion propensity, suffices to account for the hyperinvasive doublet phenotype. This work highlights bacterial cell length heterogeneity as a key determinant of target search patterns atop epithelia.

Microbial synthesis and evaluation of bactericidal properties of cadmium sulfide nanoparticles

Introduction. The productivity of microbial synthesis of stable nanoparticles is determined by the growth stage of the populations of bacterial cultures used to obtain nanostructures. The study of the biocidal activity of biogenic nanoparticles of cadmium sulfide (NPsCdS), comparable in properties with nanomaterials obtained by physicochemical methods, is promising.The aim of this work was to evaluate the effect of the cell growth phase of the bacterial strains Bacillus subtilis 168 and Shewanella oneidensis MR-1 on the efficiency of biosynthesis of NPsCdS and to study their bactericidal properties against a number of gram-positive and gram-negative strains of microorganisms.Material and methods. Nanoparticles were obtained by introducing Na2S and CdCl2 salts to a final concentration of 2 mM : 2 mM in liquid bacterial cultures with cells in different phases of growth. The efficiency of NPsCdS biosynthesis was evaluated by the optical density of aqueous nanoparticles solutions. The bactericidal properties of NPsCdS were determined by the diameter of zone of inhibition growth of gram-positive bacteria B. subtilis 168, B. amyloliquefaciens, Streptococcus salivarius, Rhodococcus rhodochrous and gram-negative S. oneidensis MR-1, Escherichia coli K-12, Pseudomonas putida.Results. It was found that the use of cells in the stationary phase of growth (18–24 hours) contributes to obtaining the maximum amount of NPsCdS corresponding to concentrations of 1.0–1.2 mg/ml. The high antimicrobial activity of NPsCdS was shown against gram-positive microorganisms, among gram-negative bacteria, P. putida strain showed insignificant sensitivity.Discussion. The experimental results expand scientific data about the effect of the phase of bacterial growth cycle on biosynthesis of nanoparticles. The stationary phase of growth of B. subtilis 168, S. oneidensis MR-1 is optimal for obtaining of NPsCdS. For the first time, the cytotoxicity of NPsCdS/Shewanella against bacteria of various taxonomic groups was demonstrated.Conclusion. An effective method for obtaining extracellular NPsCdS using bacteria B. subtilis 168, S. oneidensis MR-1 in the stationary phase of growth has been developed. The biocidal activity of biogenic NPsCdS was shown, which allows to consider them as a new class of antimicrobial agents.

DNA sequence-directed cooperation between nucleoid-associated proteins

SummaryNucleoid-associated proteins (NAPs) are a class of highly abundant DNA-binding proteins in bacteria and archaea. While both the composition and relative abundance of the NAPs change during the bacterial growth cycle, surprisingly little is known about their crosstalk in mutually binding and stabilizing higher-order nucleoprotein complexes in the bacterial chromosome. Here, we use atomic force microscopy and solid-state nanopores to investigate long-range nucleoprotein structures formed by the binding of two major NAPs, FIS and H-NS, to DNA molecules with distinct binding site arrangements. We find that spatial organization of the protein binding sites can govern the higher-order architecture of the nucleoprotein complexes. Based on sequence arrangement the complexes differed in their global shape and compaction as well as the extent of FIS and H-NS binding. Our observations highlight the important role the DNA sequence plays in driving structural differentiation within the bacterial chromosome.

Quantitative analysis of the effects of morphological changes on extracellular electron transfer rates in cyanobacteria

BackgroundUnderstanding the extracellular electron transport pathways in cyanobacteria is a major factor towards developing biophotovoltaics. Stressing cyanobacteria cells environmentally and then probing changes in physiology or metabolism following a significant change in electron transfer rates is a common approach for investigating the electron path from cell to electrode. However, such studies have not explored how the cells’ concurrent morphological adaptations to the applied stresses affect electron transfer rates. In this paper, we establish a ratio to quantify this effect in mediated systems and apply it to Synechococcus elongatus sp. PCC7942 cells grown under different nutritional regimes.ResultsThe results provide evidence that wider and longer cells with larger surface areas have faster mediated electron transfer rates. For rod-shaped cells, increase in cell area as a result of cell elongation more than compensates for the associated decline in mass transfer coefficients, resulting in faster electron transfer. In addition, the results demonstrate that the extent to which morphological adaptations account for the changes in electron transfer rates changes over the bacterial growth cycle, such that investigations probing physiological and metabolic changes are meaningful only at certain time periods.ConclusionA simple ratio for quantitatively evaluating the effects of cell morphology adaptations on electron transfer rates has been defined. Furthermore, the study points to engineering cell shape, either via environmental conditioning or genetic engineering, as a potential strategy for improving the performance of biophotovoltaic devices.

Effects of incorporating calcined corals as natural antimicrobial agent into active packaging system for milk storage.

A series of nylon (NY)/linear low-density polyethylene (LLDPE) containing calcined corals (NY/LL-CORALS) composite films were prepared using the cast extrusion method. We investigated the effect of different contents of incorporated calcined corals on the physical properties and antimicrobial activity of the composite films as well as their feasibility for milk storage applications. The results indicated that the main compound in calcined corals was calcium oxide (CaO). As the calcined corals content increased, the crystallinity of the composite films slightly decreased, but no significant changes in their thermal stability and permeability were observed. The NY/LL-CORALS composite films exhibited excellent antimicrobial performance against Escherichia coli and Staphylococcus aureus. Notably, the NY/LL-CORALS packaging significantly extended the lag time of bacteria and delayed the bacterial growth cycle in milk during storage. Thus, the NY/LL-CORALS composite films could be a potential food packaging material that could prolong the shelf life of fresh food.

Dividing subpopulation of Escherichia coli in stationary phase

The bacterial growth cycle contains different phases: after the growth substrate is exhausted or the toxic waste products accumulate the growth stops. In this non-growing culture the number of colony forming bacteria remains constant or starts to decrease. It has been shown that during prolonged incubation there is constant growth and death of bacteria and certain mutant populations take over the culture. Here we show that the dynamic cell division and death balance can be obtained even before the appearance of mutants. The possible presence of different subpopulation should be considered when analyzing physiological states of bacterial cultures.

DNA Sequence-Directed Cooperation between Nucleoid-Associated Proteins

Nucleoid associated proteins (NAPs) are a class of highly abundant DNA binding proteins in bacteria and archaea. While both the composition and relative abundance of the NAPs change during the bacterial growth cycle, surprisingly little is known about their crosstalk in mutually binding and stabilising higher-order nucleoprotein complexes in the bacterial chromosome. Here, we use atomic force microscopy and solid-state nanopores to investigate long-range nucleoprotein structures formed by the binding of two major NAPs, FIS and H-NS, to DNA molecules with distinct binding-site arrangements. We find that spatial organisation of the protein binding sites can govern the higher-order architecture of the nucleoprotein complexes. Based on sequence arrangement the complexes differed in their global shape and compaction, as well as the extent of FIS and H-NS binding. Our observations highlight the important role the DNA sequence plays in driving structural differentiation within the bacterial chromosome.

CRISPR-Cas, DNA Supercoiling, and Nucleoid-Associated Proteins.

In this opinion article we highlight links between the H-NS nucleoid-associated protein, variable DNA topology, the regulation of CRISPR-cas locus expression, CRISPR-Cas activity, and the recruitment of novel genetic information by the CRISPR array. We propose that the requirement that the invading mobile genetic element be negatively supercoiled limits effective CRISPR action to a window in the bacterial growth cycle when DNA topology is optimal, and that this same window is used for the efficient integration of new spacer sequences at the CRISPR array. H-NS silences CRISPR promoters, and we propose that antagonists of H-NS, such as the LeuO transcription factor, provide a basis for a stochastic genetic switch that acts at random in each cell in the bacterial population. In addition, we wish to propose a mechanism by which mobile genetic elements can suppress CRISPR-cas transcription using H-NS homologues. Although the individual components of this network are known, we propose a new model in which they are integrated and linked to the physiological state of the bacterium. The model provides a basis for cell-to-cell variation in the expression and performance of CRISPR systems in bacterial populations.

Evaluating the level of nitroreductase activity in clinical Klebsiella pneumoniae isolates to support strategies for nitro drug and prodrug development

To understand the potential utility of novel nitroreductase (NR)-activated prodrugs, NR enzyme activity was assessed in clinical Klebsiella pneumoniae isolates using a NR-activated fluorescent probe. NR activity was constant throughout the bacterial growth cycle, but individual K. pneumoniae isolates exhibited a wide range of NR activity levels. The genes of major NR enzymes (nfsA and nfnB) showed a number of sequence variants. Aside from a C-terminal extension of NfnB, which may be responsible for lower NR activity in specific isolates, the genetic differences did not explain the variation in activity. Analysis of important clinical strains (ST11, ST258, ST14 and ST101) showed significant variation in NR activity between isolates within the same sequence type despite conservation of nfsA/nfnB sequences. Addition of methyl viologen (MV), a known activator of soxRS, caused a significant increase in NR activity for all strains, with proportionally larger increases in activity seen for strains with low uninduced NR levels. Real-time PCR on selected strains following exposure to MV showed upregulation of soxS (15-32-fold) and nfsA (5-22-fold) in all strains tested. Expression of nfnB was upregulated 2-5-fold in 4/6 strains tested. High levels of NR activity in the absence of MV activation correlated with nitrofurantoin susceptibility. These data provide evidence that NR gene mutations and regulatory pathways influence NR activity in K. pneumoniae isolates and this is likely to impact treatment efficacy with novel nitro-containing drugs or prodrugs.

Elucidating the functional role of Mycobacterium smegmatis recX in stress response

The RecX protein has attracted considerable interest because the recX mutants exhibit multiple phenotypes associated with RecA functions. To further our understanding of the functional relationship between recA and recX, the effect of different stress treatments on their expression profiles, cell yield and viability were investigated. A significant correlation was found between the expression of Mycobacterium smegmatis recA and recX genes at different stages of growth, and in response to different stress treatments albeit recX exhibiting lower transcript and protein abundance at the mid-log and stationary phases of the bacterial growth cycle. To ascertain their roles in vivo, a targeted deletion of the recX and recArecX was performed in M. smegmatis. The growth kinetics of these mutant strains and their sensitivity patterns to different stress treatments were assessed relative to the wild-type strain. The deletion of recA affected normal cell growth and survival, while recX deletion showed no significant effect. Interestingly, deletion of both recX and recA genes results in a phenotype that is intermediate between the phenotypes of the ΔrecA mutant and the wild-type strain. Collectively, these results reveal a previously unrecognized role for M. smegmatis recX and support the notion that it may regulate a subset of the yet unknown genes involved in normal cell growth and DNA-damage repair.

A comprehensive genomic and growth proteomic analysis of antitumor lipopeptide bacillomycin Lb biosynthesis in Bacillus amyloliquefaciens X030.

Lipopeptides (such as iturin, fengycin, and surfactin) from Bacillus possess antibacterial, antifungal, and antiviral activities and have important application in agriculture and pharmaceuticals. Although unremitting efforts have been devoted to improve lipopeptide production by designing gene regulatory circuits or optimizing fermentation process, little attention has been paid to utilizing multi-omics for systematically mining core genes and proteins during the bacterial growth cycle. Here, lipopeptide bacillomycin Lb from new Bacillus amyloliquefaciens X030 was isolated and first found to have anticancer activity in various cancer cells (such as SMMC-7721 and MDA-MB-231). A comprehensive genomic and growth proteomic analysis of X030 revealed bacillomycin Lb biosynthetic gene cluster, key enzymes and potential regulatory proteins (PerR, PhoP, CcpA, and CsfB), and novel links between primary metabolism and bacillomycin Lb production in X030. The antitumor activity of the fermentation supernatant supplemented with amino acids (such as glutamic acid) and sucrose was significantly increased, verifying the role of key metabolic switches in the metabolic regulatory network. Quantitative real-time PCR analysis confirmed that 7 differential expressed genes exhibited a positive correlation between changes at transcriptional and translational levels. The study not only will stimulate the deeper and wider antitumor study of lipopeptides but also provide a comprehensive database, which promotes an in-depth analysis of pathways and networks for complex events in lipopeptide biosynthesis and regulation and gives great help in improving the yield of bacillomycin Lb (media optimization, genetic modification, or pathway engineering).

Prediction of different antibacterial activity in a new set of formyl hydroxyamino derivatives with potent action on peptide deformylase using structural information

Due to the essential role of peptide deformylase (PDF) at the bacterial growth cycle, it is a noteworthy target for developing a novel antibacterial agent. In the current study, the antibacterial activities of a set of 44 new structures of formyl hydroxyamino derivatives as PDF inhibitors were quantified using quantitative structure–activity relationship (QSAR). Artificial neural networks (ANN) were used as a chemometrics tool for QSAR modeling. Three quantitative models were suggested to relate the chemical structural features of the formyl hydroxyamino derivatives to their antibacterial activities (pIC50) against Staphylococcus aureus, methicillin-susceptible S. aureus (MSSA), and methicillin-resistant S. aureus (MRSA) peptide deformylase. The sufficiency of the model for prediction of the antibacterial activities of the desired PDF inhibitor compounds against S. aureus, MSSA, and MRSA was statistically demonstrated according to the validation parameters such as coefficient of determination (R2), mean square error (MSE) in training, validation, and prediction sets, and also using applicability domain (AD) and randomization test.

Bacterial communication systems: A mathematical formulation of negative chemotaxis

SummaryThe emergence of molecular communication (MC) has provided a new paradigm for communication at nanoscale, besides the electromagnetic one. Numerous biological systems have been proposed that could be used for communication, using mostly bacteria as information carriers, ensuring biocompatibility and low complexity requirements. In this work, message dissemination dynamics in a bacterial communication system was under investigation by means of a simulation framework that was designed and developed based on a commercial tool. A mathematical formulation is proposed, in an effort to accurately predict the system's behavior under the presence of chemotactic‐like nodes. Simulation results are in strong agreement with the respective mathematical model, for a wide range of biological processes, including phenomena such as chemotaxis, bacterial growth cycle, and biological message transformation dynamics. In the case of a simple simulation scenario exhibiting chemotaxis, the mean accuracy improvement reaches values of up to 19%.

Classical and Bayesian predictions applied to Bacillus toxin production.

Bacillus thuringiensis is a bacterium with unusual properties that make it useful for pest control in ecoagriculture. It can form a parasporal crystal containing polypeptides (also called delta-endotoxins). These entomopathogenic toxins are made during the stationary phase of the bacterial growth cycle and were initially characterized as an insect pathogen. Nowadays, the use of saturated two-level designs is very popular. This method is especially used in industrial applications where the cost of experiments is expensive. Standard classical approaches are not appropriate to analyze data from saturated designs. It is due to the fact that they only allow to estimate the main factor effects and cannot assure enough freedom degrees to estimate the error variance. In this paper, we propose the use of empirical Bayesian procedures to get inferences for data obtained from saturated designs, inspired from Hadamard matrices. The proposed methodology is illustrated by assuming a dataset to prove the model robustness. The comparison between the two studied mathematical techniques, based on mean square error values (MSE), revealed that Bayesian method (BM) was more accurate than least square method (LSM): for example, the results showed that 2002 and 2000.7 mg/l for experimental and predicted (BM) data were obtained against 2002 and 1991 mg/l for experimental and predicted (LSM) data. This suggested method could be generalized in several application fields in biological sciences.

Bacterial N2-fixation in mangrove ecosystems: insights from a diazotroph-mangrove interaction.

Mangrove forests are highly productive ecosystems but represent low nutrient environments. Nitrogen availability is one of the main factors limiting mangrove growth. Diazotrophs have been identified as key organisms that provide nitrogen to these environments. N2-fixation by such organisms was found to be higher in the mangrove roots than in surrounding rhizosphere. Moreover, previous studies showed that mangroves grew better in the presence of N2-fixers indicating a potentially mutualistic relationship. However, the molecular signals and mechanisms that govern these interactions are still poorly understood. Here we present novel insights in the interaction of a diazotroph with a mangrove species to improve our understanding of the molecular and ecophysiological relationship between these two organisms under controlled conditions. Our results showed that Marinobacterium mangrovicola is a versatile organism capable of competing with other organisms to survive for long periods in mangrove soils. N2-fixation by this bacterium was up-regulated in the presence of mangrove roots, indicating a possible beneficial interaction. The increase in N2-fixation was limited to cells of the exponential growth phase suggesting that N2-fixation differs over the bacterial growth cycle. Bacterial transformants harboring a transcriptional nifH::gusA fusion showed that M. mangrovicola successfully colonized mangrove roots and simultaneously conducted N2-fixation. The colonization process was stimulated by the lack of an external carbon source suggesting a possible mutualistic relationship. M. mangrovicola represents an interesting genetically accessible diazotroph, which colonize mangrove roots and exhibit higher N2-fixation in the presence of mangrove roots. Consequently, we propose this microorganism as a tool to study molecular interactions between N2-fixers and mangrove plants and to better understand how changes in the environment could impact these important and relatively unknown interactions.