, 2004; Hofemeister et al., 2004; López et al., 2009). ComX is a quorum-sensing peptide pheromone that triggers the production of surfactin. The lipopeptide is then involved in a paracrine signaling pathway that triggers a subpopulation of

cells to produce an extracellular matrix. Interestingly, the surfactin-producing cells do not produce a matrix themselves, but upstream activation of comX is needed for biofilm production (Magnuson et al., 1994; López et al., 2009). It is still unclear how ComX-producing cells activate surfactin synthesis and how surfactin can then trigger matrix production. In B. subtilis buy PD0325901 168 strains, single-base duplications in sfp genes cause impairment in surfactin production (Zeigler et al., 2008). This mutation also PARP inhibitor produces losses of swarming and affects the speed of colonization (Julkowska et al., 2005). sfp encodes a phosphopantetheinyl transferase that activates the peptidyl carrier protein domain of the first three subunits (SrfABC) of surfactin synthetase (Quadri et al., 1998). Microorganisms, which require the activation of carrier

proteins involved in secondary metabolic pathways, such as nonribosomal peptide synthetase or polyketide synthase pathways, require the activity of these Sfp-like proteins (Copp et al., 2007). Consequently, in the absence of the Sfp enzyme, B. subtilis cannot synthesize compounds such as surfactin, Abiraterone chemical structure which are dependent on nonribosomal peptide synthetase or polyketide synthase-type mechanisms. Bacillus subtilis strain 3610 that carries the intact sfp gene swarms rapidly in symmetrical concentric waves, forming branched dendritic

patterns. This observation was confirmed by Debois et al. (2008), who reported that surfactin molecules with a specific chain length play an important role in the swarming of communities on the agar surface. Although the specific mechanisms of surfactant secretion are unknown, lipopeptide secretion provides a powerful competitive advantage for any species during surface colonization and during competition for resources (Ron & Rosenberg, 2001). For example, surfactin produced by B. subtilis inhibits Streptomyces coelicolor aerial development and causes altered expression of developmental genes (Straight et al., 2006). It has also been established that surfactin is required for the formation of aerial structures on B. subtilis biofilm (Branda et al., 2001). The ecological role of the aerial structures is to increase the spore dispersal capacity. The second and third groups of surfactants produced by B. subtilis are peptides belonging to the iturin and plipastatin–fengycin groups, respectively (Fig. 1). Using HPLC, Ahimou et al. (2000) reported considerable variations in the lipopeptide content of seven B. subtilis strains. Among the three types of lipopeptides, only iturin A was produced by all seven B. subtilis strains.

, 2006). Reaction mixture I (50 μL) contained 100 mM HEPES (pH 7.0), 10 mM α-ketoglutarate, 0.5 mM FeSO4·7H2O, 0.5 mM ascorbate, variable concentrations (0.3–40 mM) of l-leucine, l-threonine or l-methionine, and aliquots of purified dioxygenase. Reaction I was incubated at 30 °C for 30 min, at which point it was arrested by placement on ice. The amount of enzyme applied

was selected to ensure that the increase in synthesized succinate was linear beta-catenin activation during the reaction. To determine the concentration of the synthesized succinate, 2.5 μL of reaction mixture I was added to reaction mixture II (up to final volume of 25 μL), which contained 100 mM Tris–HCl (pH 8.0), 1 mM phosphoenolpyruvate, 0.3 mM NADH, 10 mM MgCl2, 0.3 mM CoA, 0.3 mM ATP, 3 μg succinyl-coenzyme A synthetase Rucaparib purchase E. coli (purified by IMAC as his6-tag-fused protein) and 0.25 μL of a solution of pyruvate kinase (PK)/lactate dehydrogenase (LDH) from rabbit muscle (Sigma) (0.186 U of PK and 0.226 U of LDH). Reaction II was incubated at 30 °C for 1 h and halted by placement on ice. Subsequently, the absorbance at 340 nm was measured, and the concentration of synthesized succinate was deduced from a calibration curve obtained by performing reaction

II with succinate standards. Enzymatic activity was quantified by measuring the amount of succinate produced per minute and per milligram of enzyme. The KM and Vmax parameters with standard errors for l-leucine, l-threonine and l-methionine were deduced from Michaelis–Menten kinetic equation plots obtained from nonlinear regression analysis of experimental data using SigmaPlot (http://www.systat.com). The preparation and identification of l-methionine sulfoxide and hydroxylated l-leucine was performed as previously described (Hibi et al., 2011). A biomass sample of BL21(DE3) [pET-HT-BPE] strain from fresh-made LB-agar plates was inoculated into 400 mL of LB broth (2 × 200 mL) supplemented with Ap

(100 mg L−1) and cultivated at 37 °C until A555 nm = 1 was reached. Subsequently, IPTG was added to a final concentration of 1 mM, and the culture was incubated for an additional 2 h. The biomass was harvested by centrifugation and re-suspended in 5 mL of 50 mM HEPES (pH 7) and lysed by one pass through a French press (1000 psi). The five reaction mixtures (2 mL volume) Methocarbamol then consisted of 25 mM l-threonine, 25 mM α-ketoglutarate, 100 mM HEPES (pH 7), 10 mM FeSO4·7H2O and 1 mL of cell lysate. The reactions were incubated at 37 °C for 15 h with vigorous shaking. Amino acid hydroxylation was monitored by TLC analysis using ninhydrin (2-propanol/acetone/ammonia/water = 25 : 25 : 6 : 4). A 10 mL volume of the resulting supernatant was passed through a 0.22 μm filter and applied to a preparative TLC plate. The hydroxylated l-Thr was collected, eluted with water, freeze-dried and analysed by ESI-MS as described in (Hibi et al., 2011).

This research was supported by the South Transdanubian Regional Knowledge Centre (RET-08/2005, OMFB-00846/2005) and Iparjog 08 (NKTH IPARJOG-08-1-2009-0026). “
“Vibrio fischeri induces both

anaerobic respiration and bioluminescence during symbiotic infection. In many bacteria, the oxygen-sensitive regulator FNR activates anaerobic respiration, and a preliminary study using the light-generating lux genes from V. fischeri MJ1 cloned in Escherichia coli suggested that FNR stimulates bioluminescence. To test for FNR-mediated regulation of bioluminescence and anaerobic respiration in V. fischeri, we generated fnr mutants of V. fischeri strains MJ1 and ES114. In both strains, FNR was required for normal fumarate- and nitrate-dependent RG7420 clinical trial respiration. However, contrary to the report in transgenic E. coli, FNR mediated the repression of lux. ArcA represses bioluminescence, and ParcA-lacZ reporters showed reduced expression in fnr mutants, suggesting a possible indirect effect of FNR on bioluminescence via arcA. Finally, the fnr mutant of ES114 was not impaired in colonization of its host squid, Euprymna scolopes. This study extends the characterization

of FNR to the Vibrionaceae and underscores the importance of studying lux regulation in its native background. Vibrio fischeri is a model for investigations of bioluminescence and mutualistic symbioses, two fields connected by the importance of oxygen. O2 is learn more a substrate for the luminescence-producing

enzyme luciferase, and luciferase may benefit V. fischeri by generating Protirelin a more reduced environment in or near cells (Visick et al., 2000; Timmins et al., 2001). Reduction of O2 could be especially advantageous for this facultative anaerobe when it is colonizing animal tissue and may minimize the host’s ability to generate reactive oxygen species (Visick et al., 2000). Luminescence emanating from bacteria colonizing the symbiotic light organ of the host indicates that O2 is present; however, evidence suggests that luciferase is O2 limited in this environment (Boettcher et al., 1996) despite its high affinity (Km∼35 nM) for O2 (Bourgois et al., 2001). Moreover, anaerobic respiration is apparently induced in symbiotic V. fischeri (Proctor & Gunsalus, 2000), consistent with the idea that [O2] is low in the light organ. One regulator that might control anaerobic respiration and luminescence in response to [O2] is FNR (so named for its role in fumarate and nitrate reduction). FNR regulates genes during the switch between aerobic and anaerobic growth in Escherichia coli and other bacteria, and it often activates genes responsible for anaerobic respiration (Browning et al., 2002; Reents et al., 2006; Fink et al., 2007).

Mechanistic investigations revealed that the neuronal and behavioral recovery produced by

exercise in the chronic parkinsonian mice was associated with an improved mitochondrial function and an increase in the brain region-specific levels of brain-derived and glial cell line-derived neurotrophic factors. Our findings indicate that exercise not only produces neuronal and mitochondrial protection, NVP-AUY922 it also boosts nigrostriatal neurotrophic factor levels in the chronic parkinsonian mice with moderate neurodegeneration. Therefore, modifying lifestyle with increased exercise activity would be a non-pharmacological neuroprotective approach for averting neurodegenerative processes, as demonstrated in experimental chronic parkinsonism. “
“A key feature of early visual cortical regions is that they contain

discretely organized retinotopic maps. Titration of these maps must occur through experience, and the fidelity of their spatial tuning will depend on the consistency and accuracy of the eye movement system. Anomalies in fixation patterns and the ballistics of eye movements are well documented in autism spectrum disorder (ASD), with off-center fixations a hallmark of the phenotype. We hypothesized that these atypicalities might affect the development of visuo-spatial maps and specifically that peripheral inputs might receive altered processing in ASD. Using high-density recordings of visual evoked potentials PD-0332991 concentration (VEPs) and a novel system-identification approach known as VESPA (visual evoked spread spectrum analysis), we assessed sensory responses to centrally and peripherally presented stimuli. Additionally, input luminance was varied to bias

responsiveness to the magnocellular system, given previous suggestions of magnocellular-specific deficits in ASD. Participants were 22 ASD children (7–17 years of age) and 31 age- and performance-IQ-matched neurotypical controls. Both VEP and VESPA responses to central presentations were indistinguishable between groups. In contrast, peripheral presentations resulted in significantly greater early VEP and VESPA amplitudes in the ASD cohort. We found no evidence that anomalous enhancement was restricted to magnocellular-biased responses. The extent of peripheral response enhancement was related Thymidine kinase to the severity of stereotyped behaviors and restricted interests, cardinal symptoms of ASD. The current results point to differential visuo-spatial cortical mapping in ASD, shedding light on the consequences of peculiarities in gaze and stereotyped visual behaviors often reported by clinicians working with this population. Atypicalities in how individuals with an autism spectrum disorder (ASD) direct their gaze to socially relevant stimuli such as faces and eyes have long been noted (Klin et al., 2002; Pelphrey et al., 2002; Hernandez et al., 2009; Kliemann et al., 2010).

This suggests that the alterations of virB may differ from that of vjbR in some aspects.

To survive in host cells, intracellular bacteria check details have developed the capability to adapt to intracellular environments. The intracellular hostile environments include oxidative burst, high salt and high osmosis. BMΔvirB showed reduced survival capability under the stress conditions compared with BM and BM-IVGT. Sensitivity to high salt and osmosis is closely related to OM properties. Therefore, it is possible that the increased sensitivity of the virB mutant results from a modified OM structure. The T4SS is a membrane-associated structure that has been identified in a variety of

bacterial species and has multiple functions. One function of T4SS of Brucella is to direct intracellular trafficking of BCV to reach a replication niche in the ER. During this process, effector proteins may play essential roles. A recent study showed that two proteins, VceA and VceC, were translocated by T4SS into a macrophage (de Jong et al., 2008). It is possible that the two effectors, as well as other unidentified effector proteins, are involved in the virB-mediated intracellular survival of Brucella (Zhong et al., 2009). In this study, we analyzed the effect of T4SS on the OM properties Ribociclib manufacturer of B. melitensis. On the one hand, comparative proteomics and qRT-PCR revealed that T4SS affects the expression of Omp25/Omp31

and other OMPs, and that the virB mutant has a higher susceptibility to the environmental stresses. On the other hand, clumping phenotype and susceptibility assays confirmed that the virB mutant displayed altered OM properties. Therefore, in addition to effector secretion, as a membrane structure, T4SS also affects the expression of major OMPs and the properties of the OM, possibly promoting the adaptation of Brucella to environments and being indirectly related to bacterial survival. This work was supported RVX-208 by the National Natural Science Foundation of China (Grant No. 30600024) and the National High Technology Research and Development Program of China (Grant No. 2007AA02Z412). Y.W. and Z.C. contributed equally to this work. “
“Aspergillus flavus is one of the most common contaminants that produces aflatoxins in foodstuffs. It is also a human allergen and a pathogen of animals and plants. Aspergillus flavus is included in the Aspergillus section Flavi that comprises 11 closely related species producing different profiles of secondary metabolites. A six-step strategy has been developed that allows identification of nine of the 11 species. First, three real-time PCR reactions allowed us to discriminate four groups within the section: (1) A.

In this report, we present microscopic-based evidences that the TIMM process actually starts with a septation defect, leading to aberrant cell morphologies. Moreover, the septation defect of CH34 could be induced by NaOCl, thus showing that the TIMM phenotype may be part of a more general stress response. Sequence analysis of a TIMM survivor exhibiting a recurrent recognizable

lysA mutation ruled out the possibility of FDA-approved Drug Library a genetic ground linking TIMM survival and peptidoglycan synthesis. “
“Luminous marine bacteria usually emit bluish-green light with a peak emission wavelength (λmax) at about 490 nm. Some species belonging to the genus Photobacterium are exceptions, producing an accessory blue fluorescent protein (lumazine protein: LumP) that causes a blue shift, from λmax ≈ 490 to λmax ≈ 476 nm. However, the incidence of blue-shifted light emission or the

presence of accessory fluorescent proteins in bacteria of the genus Vibrio has never been reported. From our spectral analysis of light emitted by 16 luminous strains of the genus Vibrio, it was revealed that most strains of Vibrio azureus emit a blue-shifted light with a peak at approximately 472 nm, whereas other Vibrio strains emit light with a peak at around 482 nm. Therefore, we investigated the mechanism underlying this blue shift in V. azureus NBRC 104587T. Here, we describe the blue-shifted light emission spectra and the isolation of a blue fluorescent protein. Intracellular protein analyses showed that this strain had a blue fluorescent find more protein (that we termed VA-BFP), the fluorescent spectrum of which was

Nintedanib (BIBF 1120) almost identical to that of the in vivo light emission spectrum of the strain. This result strongly suggested that VA-BFP was responsible for the blue-shifted light emission of V. azureus. Luminous bacteria occur ubiquitously in marine environments and have been isolated from seawater, sediment, detritus, and light-emitting organs of marine animals (Reichelt & Baumann, 1973; Ramesh et al., 1990; Nealson & Hastings, 1991; Dunlap & Kita-Tsukamoto, 2006). To date, 23 species of luminous marine bacteria have been identified, consisting of 11 Vibrio species, four Aliivibrio species, six Photobacterium species, and two Shewanella species (Gomez-Gil et al., 2004; Dunlap & Kita-Tsukamoto, 2006; Ast et al., 2007; Urbanczyk et al., 2007, 2008; Yoshizawa et al., 2009a, b, 2010a, b, in press). Luminous bacteria use bacterial luciferase to produce a bluish-green light. The luciferase catalyzes the oxidation of reduced riboflavin-5′-phosphate (FMNH2) with a long-chain aliphatic aldehyde and molecular oxygen, and the peak light emission generally occurs around 490 nm (Hastings & Nealson, 1977).

The characterization of the genomic variation is fundamental to understand the evolution of M. tuberculosis, its adaptation to human populations and to the immune response elicited by its host. Recent evidence has shown that M. tuberculosis genotype influences clinical disease phenotype, and that a significant interaction exists between host and bacterial genotypes for the development of tuberculosis (Nahid et al., 2010). In this report, we describe the genome characteristics of the Colombian clinical isolate UT205, which was isolated

from a patient with TB from Medellin, Antioquia. A comparison was carried out against the H37Rv reference genome. At the predicted protein level, we found changes in at least one amino acid in 430 coding sequences. Genomic differences are owing to indel events

and substitutions. One of the PI3K inhibitor most striking genomic modifications involves a 3.6 kbp deletion that ends with the loss of four genes, Selleckchem U0126 two belonging to the dosR regulon. Mycobacterium tuberculosis UT205 was isolated from sputum of a 33-year-old man with recently diagnosed tuberculosis. A single colony from Dubos solid medium was transferred to 7H9 liquid medium supplemented with OADC and Tween-80, cultured to an OD600 nm of 0.5, harvested by centrifugation and resuspended in TE pH8.0 [0.01 M Tris–HCl, 0.001 M EDTA (pH 8.0)]. For genomic DNA extraction, mycobacteria were freeze-thawed in ethanol-dry ice, heated at 80 °C, digested with lysozyme and incubated 1 h with 10% SDS at 60 °C, and again submitted to five cycles of freeze-thawing. Genomic DNA was phenol/chloroform/isoamyl alcohol (25 : 24 : 1, v/v) extracted, precipitated with isopropanol, washed with 75% ethanol and finally resuspended in TE pH8.0. Molecular characterization by IS6110 RFLP and spoligotyping (van Embden et al., 1993; Kamerbeek et al., 1997) identified this isolate as belonging to the LAM09 family after comparison selleck monoclonal humanized antibody with the sitvit2 database (Pasteur Institute of Guadeloupe). Whole genome shotgun sequencing was carried out using the ROCHE 454-GS-FLX TITANIUM technology at the National Center for Genomic Sequencing-CNSG (Medellin-Colombia), following standard

protocols. The genome assembly process was performed using the newbler v2.3 software with default settings. Contig reordering and joining were carried out with the ABACAS script from the Sanger institute (Assefa et al., 2009) based on the H37Rv reference genome (EMBL accession number AL123456). For genome annotation, a single fasta file containing all contigs ordered with the mummer package v3 (Delcher et al., 2003) based on the H37Rv reference genome (EMBL accession number AL123456) was built and annotated using the RATT tool from the SANGER institute (Otto et al., 2011), which transfers the genome-annotated features of a reference genome. Manual curation of the annotation was carried out with the artemis software (Rutherford et al., 2000).

The characterization of the genomic variation is fundamental to understand the evolution of M. tuberculosis, its adaptation to human populations and to the immune response elicited by its host. Recent evidence has shown that M. tuberculosis genotype influences clinical disease phenotype, and that a significant interaction exists between host and bacterial genotypes for the development of tuberculosis (Nahid et al., 2010). In this report, we describe the genome characteristics of the Colombian clinical isolate UT205, which was isolated

from a patient with TB from Medellin, Antioquia. A comparison was carried out against the H37Rv reference genome. At the predicted protein level, we found changes in at least one amino acid in 430 coding sequences. Genomic differences are owing to indel events

and substitutions. One of the Talazoparib in vitro most striking genomic modifications involves a 3.6 kbp deletion that ends with the loss of four genes, selleck two belonging to the dosR regulon. Mycobacterium tuberculosis UT205 was isolated from sputum of a 33-year-old man with recently diagnosed tuberculosis. A single colony from Dubos solid medium was transferred to 7H9 liquid medium supplemented with OADC and Tween-80, cultured to an OD600 nm of 0.5, harvested by centrifugation and resuspended in TE pH8.0 [0.01 M Tris–HCl, 0.001 M EDTA (pH 8.0)]. For genomic DNA extraction, mycobacteria were freeze-thawed in ethanol-dry ice, heated at 80 °C, digested with lysozyme and incubated 1 h with 10% SDS at 60 °C, and again submitted to five cycles of freeze-thawing. Genomic DNA was phenol/chloroform/isoamyl alcohol (25 : 24 : 1, v/v) extracted, precipitated with isopropanol, washed with 75% ethanol and finally resuspended in TE pH8.0. Molecular characterization by IS6110 RFLP and spoligotyping (van Embden et al., 1993; Kamerbeek et al., 1997) identified this isolate as belonging to the LAM09 family after comparison PtdIns(3,4)P2 with the sitvit2 database (Pasteur Institute of Guadeloupe). Whole genome shotgun sequencing was carried out using the ROCHE 454-GS-FLX TITANIUM technology at the National Center for Genomic Sequencing-CNSG (Medellin-Colombia), following standard

protocols. The genome assembly process was performed using the newbler v2.3 software with default settings. Contig reordering and joining were carried out with the ABACAS script from the Sanger institute (Assefa et al., 2009) based on the H37Rv reference genome (EMBL accession number AL123456). For genome annotation, a single fasta file containing all contigs ordered with the mummer package v3 (Delcher et al., 2003) based on the H37Rv reference genome (EMBL accession number AL123456) was built and annotated using the RATT tool from the SANGER institute (Otto et al., 2011), which transfers the genome-annotated features of a reference genome. Manual curation of the annotation was carried out with the artemis software (Rutherford et al., 2000).

The relative amount of these localizations varied between experiments. As a control of free

cytoplasmic GFP, a culture that has been previously reported to produce GFP in all cells of N. punctiforme was used (Fig. 3b) (Huang et al., 2010). This GFP control culture produced homogeneously distributed GFP in the cytoplasm of the heterocysts. In the cytoplasm of vegetative cells, the fluorescence was clearly obstructed by the presence of thylakoid membranes. The presence of thylakoids is seen in the red autofluorescence (magenta) stemming from the thylakoid-attached phycobilisome/photosystem II complexes (Cardona et al., 2009) and in the limited overlap between autofluorescence and GFP fluorescence (Fig. 3b). The full-length HupS–GFP protein required strong denaturing Quizartinib mouse conditions (2% SDS) for efficient

extraction (Fig. 1b), whereas most of the HupS–GFP degradation products could be extracted without detergent (Fig. S1). To examine the potential formation of a complete uptake hydrogenase by HupS–GFP and HupL, efforts were made to prepare native DAPT research buy extracts of these proteins from N2-fixing cultures of SHG. However, none of these attempts were successful. To examine the solubility of HupS–GFP, anti-GFP Western blots were performed with proteins extracted using buffers containing no detergents, mild nonionic detergents, or strongly denaturing additives. The results show that HupS–GFP could only be efficiently extracted using the strongly denaturing additives (Fig. 1b and Fig. S1). To identify any cell structure differences caused by potential HupS–GFP protein inclusions, isolated heterocysts

from N2-fixing Non-specific serine/threonine protein kinase cultures of SHG and WT were compared using TEM. The resulting images did not reveal any structural differences between SHG and WT heterocysts (Fig. S2). This study shows that the small subunit of the uptake hydrogenase, HupS, in N. punctiforme is solely localized to the heterocysts. The localization of the uptake hydrogenase in N. punctiforme has been under debate for a long time since previous immunolocalization studies have identified the large subunit, HupL, in both vegetative cells and heterocysts (Lindblad & Sellstedt, 1990; Tamagnini et al., 2002, 2007; Seabra et al., 2009). Interestingly, these studies used several different HupL antibodies and the results are not fully correlating. Both Seabra et al. (2009) and Lindblad & Sellstedt (1990) show vegetative cell localization of HupL, but the subcellular localization to the cytoplasmic membranes between vegetative cells clearly seen in Lindblad & Sellstedt (1990) is missing in Seabra et al. (2009), which argued for a subcellular localization of an inactive form to the vegetative cell thylakoid membranes as well as to what is described as the vesicular region of the heterocysts (Seabra et al., 2009).

Cultures were grown in photoheterotrophic conditions for 45 h, at which point they are ~35 h into the stationary phase of growth. These cultures were filtered using 0.45-μm PVDF syringe filters and filtrates assayed for RcGTA activity by mixing 0.1 mL of filtrate with DW5 cells in a total volume of 0.6 mL GTA buffer (Solioz et al., 1975). After incubation for 1 h, 0.9 mL of RCV broth was added and the mixtures incubated for an additional 4 h

with shaking at 200 r.p.m. The samples were plated on YPS agar, incubated in anaerobic phototrophic conditions to select for transfer of the puhA marker, and colony numbers were counted after 48 h. RcGTA activity was calculated as a ratio relative to paired wild-type RcGTA activity in three replicate experiments. Statistically significant differences in this website RcGTA activities were identified by one-way analysis of variance (anova) in R (Chambers et al., 1993). Western blots targeting the RcGTA major capsid protein (~32 kDa) were performed on the same cultures PXD101 datasheet used for RcGTA activity assays. For each culture, 0.5 mL of culture was centrifuged at > 13 000 g for 1 min to pellet the cells, and 0.4 mL of the resulting supernatants was carefully collected into a separate tube. The cell pellets were resuspended in 0.5 mL of TE buffer. These samples, 5 μL of cells and 10 μL of supernatants, were mixed with 3× SDS–PAGE

sample buffer, boiled for 5 min at 98 °C, and run on a 10% SDS–PAGE gel. Proteins were transferred to a nitrocellulose membrane by electroblotting in transfer buffer [48 mM Tris Base, 39 mM glycine, 20% methanol (v/v)]. The presence of equivalent total protein levels within supernatant and cell sample groups was verified

by staining the blotted membrane with Ponceau-S. The membranes were rinsed and blocked with a 5% (w/v) skim milk solution in TBST [20 mM Tris, 137 mM NaCl, 0.1% Tween-20 (v/v); pH 7.5] for 1 h. The membranes were rinsed with TBST and incubated overnight at 4 °C with a primary antibody ID-8 (1 : 1000 dilution in TBST) specific for the RcGTA major capsid protein (Agrisera, Sweden) (Fu et al., 2010). The membranes were washed three times in TBST, for 5 min each, and incubated with peroxidase-conjugated anti-rabbit IgG (Santa Cruz Biotechnology) (1 : 5000 dilution in TBST) for 1 h at room temperature. The membranes were rinsed three times with TBST for 5 min each, and bands detected by chemiluminescence using the SuperSignal West Femto Reagent Kit (Thermo Fisher Scientific, Canada). Images were captured on an Alpha Innotech U400 camera and then inverted and adjusted for brightness and contrast with image processing software. Motility assay tubes (Krieg & Gerhardt, 1981) were made with 0.35% agar YPS, and the stabs were incubated phototrophically at 35 °C. Tubes were photographed after 2 days of growth and the images adjusted for brightness and contrast with image processing software.