pseudomonas fluorescens flagella

Furthermore, by using an anti FliC1 antiserum, we have shown that the kinB mutant does not produce more FliC1 than the wild-type strain. Opsonophagocytic assay.An opsonophagocytic assay was used as previously described (2, 18) with the following modifications: to prepare bacteria for use in the assay, 3 ml instead of 10 ml of TSB was inoculated with bacteria from a tryptic soy agar (TSA) plate grown overnight at 37°C, with the inoculum placed in the TSB tubes, and bacteria were grown statically at 37°C until an OD at 650 nm (OD650) of 0.2 was obtained; RPMI medium was used with 10% heat-inactivated fetal bovine serum and 25 mM HEPES as the diluent; and infant rabbit serum adsorbed with the target strain for 1 h at 4°C was used as a complement source instead of human serum. Resis) were surface disinfected in 70% ethanol for 2 min and then in diluted bleach (1:5; final sodium hypochlorite concentration, 1%) for 15 min and rinsed thoroughly with sterile distilled water. Access scientific knowledge from anywhere. doi: 10.1046/j.1365-2958.2003.03740.x, de Lorenzo, V., Eltisb, L., Kesslerb, B., and Timmisb, K. N. (1993). Primers were designed to amplify flhDC (flhDC-F2/R2; Supplementary Table 2). Utilization of glucose and fructose by the bacteria may explain why medium from the association wheat–Azospirillum was less chemotacticaly active than medium from wheat plants alone. Wild-type bacteria showed extensive growth on tomato roots; the number of bacteria increased from 10(3) bacteria/cm of root length at the time of inoculation to more than 10(7) bacteria/cm after 10 days. DC3000. Multilocus sequence analysis of the 45 ORFs concatenated sequences from the second flagellar island shows agreement with the proposed four insertion events and the phylogenetic distance between the genomes harboring the island. At 130 days after seeding no effect of any of the seed tuber treatments was found in both short and long rotations of potato. 1), consistent with the titer determinations (Table 1). A., and O’Brien, A. D. (1994). Some groups of plant-growth-promoting rhizobacteria are involved in an indirect mechanism, either by their antagonistic effect against phytopathogens or by induced systemic resistance (ISR) mechanisms in plants. This flagellum is also produced under laboratory conditions by the kinB and algU mutants. Different letters indicate statistically significant differences (p < 0.05). Approximately 105 log-phase organisms were inoculated onto plates made with lysogeny broth (LB) and 0.3% agar in the presence or absence of various dilutions of antisera raised to flagellin or flagella and incubated at 30°C for 18 h. Results were photographically recorded after 18 h. Flagellar typing of CF strains.Flagellar typing of CF strains was accomplished by PCR amplification of the central region of the flagellin gene using primers specific for N-terminal (CW46 [5′-GGCCTGCAGATCNCCAA-3′]) and C-terminal (CW45 [5′-GGCAGCTGGTTNGCCTG-3′]) conserved regions as previously described (8, 43). In order to confirm that hyperflagellation resulted from production of the second flagellar apparatus and not by hyperproduction of the first flagellum, we extracted extracellular proteins from cultures of the wild-type strain, the kinB mutant and a sadB mutant, that has been previously shown to overproduce the FliC flagellin. Pseudomonas abietaniphila; P. agarici; P. agarolyticus; P. alcaliphila; P. alginovora; P. andersonii; P. antarctica; P. asplenii; P. azelaica; P. batumici; P. borealis; P. brassicacearum; P. chloritidismutans; P. cremoricolorata; P. diterpeniphila; P. filiscindens; P. frederiksbergensis; P. gingeri; P. graminis; P. grimo… Production of the second flagellar apparatus in F113 results in a tuft of polar flagella, which increases motility, and gives a competitive advantage for root colonization. BLAST + : architecture and applications. Seed tuber treatment with cell suspensions of fluorescent antagonistic pseudomonads in carboxymethylcellulose (CMC) raised tuber yield in potato-soil up to 70% (550 % increase), number of tubers up to 93% (934 % increase) and total plant weight up to 90% (31 % increase) as compared with CMC-treatment in wheat-soil. AlgU is required for the expression of kinB and amrZ. Over 12% of the the genes were found to deal with regulation; this may be due to the bacterium's need "for rapid adaptation to the diverse environments encountered during epiphytic growth and pathogenesis" (Buell et al. In these assays, NRS was used as a negative control. 2A), while an antiserum to type b flagellin had no killing activity (EC50 < 4) against homologous strain PA01 or heterologous strain PAK (Fig. Mutation of fliC2 in the kinB- background restored the formation of a single polar flagellum, showing that the tuft represents the second flagellar apparatus. Dialysis (6000 molecular weight cut-off) of exudate nullified its attractiveness to RW1; heating (121 °C, 15 min) of exudate had no effect. Vaccines based on monomeric flagellin have shown some efficacy in animals when a DNA vaccine encoding P. aeruginosa flagellin lacking TLR5 agonist activity is used (33), as has a multimeric construct of OprF(311-341)-OprI-flagellin fusion proteins (42) in models of burn wound and pulmonary infection. Since the phenotypic variant S appeared to be producing the second flagellar apparatus, we decided to test fliC2 expression in a battery of hypermotile phenotypic variants isolated from alfalfa rhizosphere (Martinez-Granero et al., 2006). Apparently growth-inhibiting substances other than siderophores were involved. Estimated dilution of serum needed to mediate killing of 50% (EC50) of the target bacterial strain in an opsonic killing assay. Quantification of the 16S rRNA gene confirmed the higher abundance of bacteria in the rhizosphere. We also tested the competitive colonization ability of the wild-type strain against a fliC2 mutant, unable to produce the second flagella. Microbiol. 71, 2687–2694. The second flagella is cryptic, but is produced by mutants in the kinB gene or by the wild-type strain when ectopically expressing the flhDC operon. The genomic sequence of Pseudomonas fluorescens F113 has shown the presence of a 41 kb cluster of genes that encode the production of a second flagellar apparatus. Antiserum to polymeric type a flagella was quite active, with an EC50 of 937 against homologous strain PAK (Fig. Production of a second flagellar apparatus is an unusual trait in fluorescent pseudomonads. The region also showed synteny with the flagellar genes of A. vinelandii. 56, 100–122. (A) RT-PCR of the fliC2 gene in P. fluorescens F113 and different isogenic mutants. In order to examine the spoilage of dairy products from bacteria, Dogan and Boor isolated 338 different species of Pseudomonas from raw milk, processed milk, and the environment. Pseudomonas fluorescens produces the PK antibiotic mupirocin (mup) which is active against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus.It is a mixture of pseudomonic acids, each of which comprise a C 17 monic acid (MA) and a C 9 9-hydroxynonanoic acid (9-HN) joined by an ester linkage. doi: 10.1111/j.1751-7915.2009.00103.x. Taken together, these results show that the production of the second flagella in the rhizosphere environment is an important trait for root competitive colonization. doi: 10.1146/annurev.micro.57.030502.090832, Martínez-Granero, F., Capdevila, S., Sánchez-Contreras, M., Martín, M., and Rivilla, R. (2005). doi: 10.1016/0378-1119(94)90324-7, Scher, F. M., and Baker, R. (1982). (i) Passive vaccination. Strains and plasmids used are listed in Supplementary Table 1. Tools for genetic engineering in the amino acid-producing bacterium Corynebacterium glutamicum. All animal studies were conducted in accordance with protocols approved by the Harvard Medical Area Institutional Animal Care and Use Committee. doi: 10.1128/JB.01546-10, Chin-A-Woeng, T. F., Bloemberg, G. V., Mulders, I. H., Dekkers, L. C., and Lugtenberg, B. J. The role of motility in the colonization of potato roots by Pseudomonas bacteria was studied. Join ResearchGate to find the people and research you need to help your work. Pseudomonas With untreated, tuber yield in a 1:3 (short) rotation compared to yield in a 1:6 (long) rotation of potato was decreased by 11% at 86 days (seed tuber harvest) and by 14% at 130 days (ware potato harvest) after seeding. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). After serum injections, mice were challenged i.n. The same electrophoretic conditions were used for Western blotting. Five seed-colonizing fluorescent Pseudomonas strains (RW1 to RW5) exhibited chemotaxis toward soybean seed exudates in 1-μL capillaries held for 30 min in an 8.0 log colony-forming units/mL bacterial suspension over the temperature range of 9 to 41 °C.

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