Isolation and partial characterization of a new linear DNA plasmid isolated from Bacillus polymyxa SCE2

Rosado, Alexandre Soares; Seldin, Lucy

doi:10.1099/00221287-139-6-1277

Cited by 7 publications

(5 citation statements)

References 37 publications

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“…No hybridization between pGIL01 and the linear molecules present in the type strain of B. cereus sensu stricto (ATCC 14579 T ) (Carlson et al, 1992) and in P. polymyxa (Rosado & Seldin, 1993) could be observed. Although the nature of these extrachromosomal DNA remains uncertain, it is possible that they also belong to the Tectiviridae family, which would further extend the variation spectrum of this interesting family of bacteriophages.…”

Section: Discussionmentioning

confidence: 99%

“…Linear molecules of apparently the same size as pGIL01 were also reported in the type strain of B. cereus (ATCC 14579 T ) (Carlson et al, 1992) and in strain SCE2 of Paenibacillus polymyxa (Rosado & Seldin, 1993). However, hybridization, using pGIL01 as probe, performed on these strains displayed no cross-reaction between these plasmids, suggesting that they are not closely related (data not shown).…”

Section: Uv Irradiation Mitomycin C and Nalidixic Acid Induce Gil01 mentioning

confidence: 90%

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pGIL01, a linear tectiviral plasmid prophage originating from Bacillus thuringiensis serovar israelensis

Verheust

Jensen²,

Mahillon

2003

Microbiology

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Bacillus thuringiensis serovar israelensis harbours, in addition to several circular plasmids, a small linear molecule of about 15 kb. Sequence analysis of this molecule, named pGIL01, showed the presence of at least 30 ORFs, five of which displayed similarity with proteins involved in phage systems: a B-type family DNA polymerase, a LexA-like repressor, two potential muramidases and a DNA-packaging protein (distantly related to the P9 protein of the tectiviral phage PRD1). Experimental evidence confirmed that pGIL01 indeed corresponds to the linear prophage of a temperate phage. This bacteriophage, named GIL01, produces small turbid plaques and is sensitive to organic solvents, which suggests the presence of lipid components in its capsid. Experiments using proteases and exonucleases also revealed that proteins are linked to the genomes of both pGIL01 prophage and GIL01 phage at their 59 extremities. Altogether, these features are reminiscent of those of phages found in the Tectiviridae family, and more specifically of those of PRD1, a broad-host-range phage of Gram-negative bacteria. Dot-blot hybridization, PFGE, PCR and RFLP analyses also showed the presence of pGIL01 variants in the Bacillus cereus group. INTRODUCTIONBacillus thuringiensis, the most widely used entomopathogenic bacterium, belongs to the Bacillus cereus sensu lato group. This cluster also includes B. cereus sensu stricto, an opportunist organism implicated in food poisoning (Granum & Lund, 1997), Bacillus anthracis, a human and animal pathogen, Bacillus mycoides and Bacillus pseudomycoides, characterized by their rhizoid growth (Nakamura, 1998), and the psychrotolerant Bacillus weihenstephanensis (Lechner et al., 1998). Despite their broad virulence, these bacteria are genetically closely related (their 16S RNA gene sequences share more than 99 % identity) and could be regarded as pertaining to a single species (Ash et al., 1991;Helgason et al., 2000).B. thuringiensis strains produce, during their sporulation, crystal toxins (delta-endotoxins) that are highly toxic to a number of insect larvae belonging to the orders Lepidoptera, Diptera and Coleoptera, but harmless to vertebrates. Classically, the numerous entomopathogenic B. thuringiensis strains, which have their own specific insecticidal activity, have been classified in different serotypes on the basis of their flagellar antigens. B. thuringiensis serovar (sv.) israelensis is active against dipteran species and is therefore one of the bioinsecticides of choice to control black flies and mosquitoes, both important vectors of human and animal diseases (for a recent review, see Glare & O'Callaghan, 2000).B. thuringiensis sv. israelensis strain H14 has been reported to contain at least eight DNA molecules, including three small (5?4, 6?7 and 7?6 kb) and four large (128, 145, 240 and 350 kb) circular plasmids, and one linear molecule (G. Jensen & L. Andrup, unpublished results). The pathogenicity of this strain only depends on the presence of the 128 kb plasmid which encodes the Cry and Cyt...

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Section: Discussionmentioning

confidence: 99%

Section: Uv Irradiation Mitomycin C and Nalidixic Acid Induce Gil01 mentioning

confidence: 90%

pGIL01, a linear tectiviral plasmid prophage originating from Bacillus thuringiensis serovar israelensis

Verheust

Jensen²,

Mahillon

2003

Microbiology

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“…Extraction of DNA from pure cultures and from bulk and rhizosphere soil samples. Total genomic DNA was isolated from pure cultures of bacterial strains as previously described (27). For extraction of DNA from soil, 2-g samples of bulk or rhizosphere soil were subjected to direct extraction by using a modification of the protocol of Smalla et al (36).…”

Section: Methodsmentioning

confidence: 99%

Genetic Diversity of nifH Gene Sequences in Paenibacillus azotofixans Strains and Soil Samples Analyzed by Denaturing Gradient Gel Electrophoresis of PCR-Amplified Gene Fragments

Rosado

Duarte

Seldin

et al. 1998

Appl Environ Microbiol

154

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The diversity of dinitrogenase reductase gene (nifH) fragments in Paenibacillus azotofixans strains was investigated by using molecular methods. The partial nifHgene sequences of eight P. azotofixans strains, as well as one strain each of the close relatives Paenibacillus durum,Paenibacillus polymyxa, and Paenibacillus macerans, were amplified by PCR by using degenerate primers and were characterized by DNA sequencing. We found that there are twonifH sequence clusters, designated clusters I and II, inP. azotofixans. The data further indicated that there was sequence divergence among the nifH genes of P. azotofixans strains at the DNA level. However, the gene products were more conserved at the protein level. Phylogenetic analysis showed that all nifH cluster II sequences were similar to the alternative (anf) nitrogenase sequence. A nested PCR assay for the detection of nifH (cluster I) of P. azotofixans was developed by using the degenerate primers as outer primers and two specific primers, designed on the basis of the sequence information obtained, as inner primers. The specificity of the inner primers was tested with several diazotrophic bacteria, and PCR revealed that these primers are specific for the P. azotofixans nifH gene. A GC clamp was attached to one inner primer, and a denaturing gradient gel electrophoresis (DGGE) protocol was developed to study the genetic diversity of this region of nifH inP. azotofixans strains, as well as in soil and rhizosphere samples. The results revealed sequence heterogeneity among differentnifH genes. Moreover, nifH is probably a multicopy gene in P. azotofixans. Both similarities and differences were detected in the P. azotofixans nifH DGGE profiles generated with soil and rhizosphere DNAs. The DGGE assay developed here is reproducible and provides a rapid way to assess the intraspecific genetic diversity of an important functional gene in pure cultures, as well as in environmental samples.

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“…Both strains CAPE95 and CAPE238 had their DNA isolated using the methods described by Rosado and Seldin (1993). The bacterial cultures of each strain were centrifuged (10,000 × g for 15 min), concentrated in 5 mL of Tris-EDTA-NaCl buffer (pH = 8.0), treated with 500 μL of lysozyme (10 mg mL −1 for 1 h at 37°C) and 500 μL of 10% sodium dodecyl sulfate (10 min at 37°C).…”

Section: Dna Isolation and Whole Genome Sequencingmentioning

confidence: 99%

Co-inoculation of the endophytes Bacillus thuringiensis CAPE95 and Paenibacillus polymyxa CAPE238 promotes Tropaeolum majus L. growth and enhances its root bacterial diversity

Dal’Rio,

Lopes,

Santaren

et al. 2024

Front. Microbiol.

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Tropaeolum majus L. is a versatile edible plant that is widely explored due to its medicinal properties and as a key element in intercropping systems. Its growth could be improved by the use of biofertilizers that can enhance nutrient uptake by the plant or provide tolerance to different abiotic and biotic stresses. In a previous study, 101 endophytes isolated from T. majus roots showed more than three plant growth-promoting (PGP) features in vitro, such as phosphate mineralization/solubilization, production of siderophores, antimicrobial substances and indole-related compounds, and presence of the nifH gene. To provide sustainable alternatives for biofertilization, the genomes of two promising endophytes—CAPE95 and CAPE238—were sequenced to uncover metabolic pathways related to biofertilization. Greenhouse experiments were conducted with 216 seeds and 60 seedlings, half co-inoculated with the endophytes (treatment) and half inoculated with 1X PBS (control), and the impact of the co-inoculation on the plant’s bacteriome was accessed through 16S rRNA gene metabarcoding. The strains CAPE95 and CAPE238 were taxonomically assigned as Bacillus thuringiensis and Paenibacillus polymyxa, respectively. Metabolic pathways related to the enhancement of nutrient availability (nitrogen fixation, sulfate-sulfur assimilation), biosynthesis of phytohormones (indole-3-acetic acid precursors) and antimicrobial substances (bacilysin, paenibacillin) were found in their genomes. The in vivo experiments showed that treated seeds exhibited faster germination, with a 20.3% higher germination index than the control on the eleventh day of the experiment. Additionally, treated seedlings showed significantly higher plant height and leaf diameters (p < 0.05). The bacterial community of the treated plants was significantly different from that of the control plants (p < 0.001) and showed a higher richness and diversity of species (Chao and Shannon indexes, p < 0.001). A higher relative abundance of potential synergistic PGP bacteria was also shown in the bacteriome of the treated plants, such as Lysinibacillus and Geobacter. For the first time, co-inoculation of B. thuringiensis and P. polymyxa was shown to have great potential for application as a biofertilizer to T. majus plants. The bacterial consortium used here could also be explored in other plant species in the future.

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Isolation and partial characterization of a new linear DNA plasmid isolated from Bacillus polymyxa SCE2

Cited by 7 publications

References 37 publications

pGIL01, a linear tectiviral plasmid prophage originating from Bacillus thuringiensis serovar israelensis

pGIL01, a linear tectiviral plasmid prophage originating from Bacillus thuringiensis serovar israelensis

Genetic Diversity of nifH Gene Sequences in Paenibacillus azotofixans Strains and Soil Samples Analyzed by Denaturing Gradient Gel Electrophoresis of PCR-Amplified Gene Fragments

Co-inoculation of the endophytes Bacillus thuringiensis CAPE95 and Paenibacillus polymyxa CAPE238 promotes Tropaeolum majus L. growth and enhances its root bacterial diversity

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