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An Rhs-like genetic element is involved in bacteriocin production by Pseudomonas savastanoi pv. savastanoi

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Abstract

The main aim of this work was the identification of genetic determinants involved in bacteriocin production by strain ITM317 of Pseudomonas savastanoi pv. savastanoi, besides bacteriocin characterization. The bacteriocin was observed to be a heat-sensitive, high molecular weight proteinaceous compound. We identified a transposon (Tn5)-induced mutant which had lost its ability to produce the bacteriocin. The Tn5 insertion’s responsibility for the above mutated phenotype was demonstrated by marker-exchange mutagenesis. An EcoRI DNA fragment, corresponding to the EcoRI Tn5-containing fragment of the mutant, was also cloned from the wild-type strain, and its introduction into the mutant complemented the mutation. Moreover, that fragment enabled bacteriocin production by P. s. pv. savastanoi ITM302, a strain not previously capable of doing so. DNA sequence analysis revealed that Tn5 insertion occurred in the mutant within a large ORF encoding a protein which showed similarity with proteins from the Rhs family. The DNA region including that ORF showed features which have been considered typical of the Rhs genetic elements previously identified in other bacteria but whose function is as yet unclear. The results of this study for the first time identify an Rhs-like element in P. s. pv. savastanoi, and for the first time indicate that an Rhs element is involved in bacteriocin production, also suggesting this possible function for Rhs genetic elements previously characterized in other bacteria.

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Abbreviations

Ap:

Ampicillin

Bact+ :

Able to produce the bacteriocin

Bact− :

Unable to produce the bacteriocin

Cm:

Chloramphenicol

Gm:

Gentamicin

Km:

Kanamycin

M.E.:

Marker exchange mutagenesis

r :

Resistance

Tc:

Tetracycline

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Acknowledgments

We wish to thank D. K. Willis, R. W. Jackson and J. Murillo for generously providing plasmid pRK7813, plasmid pBBR1MCS-5, and P. s. pv. tomato strain PT 23, respectively. We also thank G. Stea for his technical assistance in DNA sequencing.

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Authors and Affiliations

  1. Institute of Sciences of Food Production (ISPA), National Research Council (CNR), Via Amendola 122/O, 70126, Bari, Italy

    Angelo Sisto, Maria Morea, Stella Lisa Lonigro, Francesca Valerio & Paola Lavermicocca

  2. Plant Protection Institute, Section of Bari, National Research Council, Via Amendola 122/D, 70126, Bari, Italy

    Maria Grazia Cipriani

Authors
  1. Angelo Sisto
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  2. Maria Grazia Cipriani
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  3. Maria Morea
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  4. Stella Lisa Lonigro
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  5. Francesca Valerio
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  6. Paola Lavermicocca
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Corresponding author

Correspondence to Angelo Sisto.

Additional information

The GenBank accession number for the sequence reported in this paper is GQ259959.

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Sisto, A., Cipriani, M.G., Morea, M. et al. An Rhs-like genetic element is involved in bacteriocin production by Pseudomonas savastanoi pv. savastanoi . Antonie van Leeuwenhoek 98, 505–517 (2010). https://doi.org/10.1007/s10482-010-9468-7

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