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Population Structure and Diversity of Phenazine-1-Carboxylic Acid Producing Fluorescent Pseudomonas spp. from Dryland Cereal Fields of Central Washington State (USA)

  • Plant Microbe Interactions
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Abstract

Certain strains of the rhizosphere bacterium Pseudomonas fluorescens contain the phenazine biosynthesis operon (phzABCDEFG) and produce redox-active phenazine antibiotics that suppress a wide variety of soilborne plant pathogens. In 2007 and 2008, we isolated 412 phenazine-producing (Phz+) fluorescent Pseudomonas strains from roots of dryland wheat and barley grown in the low-precipitation region (<350 mm annual precipitation) of central Washington State. Based on results of BOX-PCR genomic fingerprinting analysis, these isolates, as well as the model biocontrol Phz+ strain P. fluorescens 2-79, were assigned to 31 distinct genotypes separated into four clusters. All of the isolates exhibited high 16S rDNA sequence similarity to members of the P. fluorescens species complex including Pseudomonas orientalis, Pseudomonas gessardii, Pseudomonas libanensis, and Pseudomonas synxantha. Further recA-based sequence analyses revealed that the majority of new Phz+ isolates (386 of 413) form a clade distinctly separated from P. fluorescens 2-79. Analysis of phzF alleles, however, revealed that the majority of those isolates (280 of 386) carried phenazine biosynthesis genes similar to those of P. fluorescens 2-79. phzF-based analyses also revealed that phenazine genes were under purifying selection and showed evidence of intracluster recombination. Phenotypic analyses using Biolog substrate utilization and observations of phenazine-1-carboxylic acid production showed considerable variability amongst members of all four clusters. Biodiversity indices indicated significant differences in diversity and evenness between the sampled sites. In summary, this study revealed a genotypically and phenotypically diverse group of phenazine producers with a population structure not seen before in indigenous rhizosphere-inhabiting Phz+ Pseudomonas spp.

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Acknowledgments

The project described was supported by award number T32GM083864 from the National Institute of General Medical Sciences. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences or the National Institutes of Health.

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Authors and Affiliations

  1. School of Molecular Biosciences, Washington State University, Pullman, WA, 99164-4234, USA

    James A. Parejko

  2. Department of Plant Pathology, Washington State University, Pullman, WA, 99164-6430, USA

    Dmitri V. Mavrodi & Olga V. Mavrodi

  3. Agricultural Research Service, USDA-ARS, Root Disease and Biological Control Research Unit, Washington State University, Pullman, WA, 99164-6430, USA

    David M. Weller & Linda S. Thomashow

  4. USDA-ARS Root Disease and Biological Control Research Unit, Washington State University, Pullman, WA, 99163-6430, USA

    Linda S. Thomashow

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  1. James A. Parejko
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  2. Dmitri V. Mavrodi
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  3. Olga V. Mavrodi
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  4. David M. Weller
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  5. Linda S. Thomashow
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Correspondence to Linda S. Thomashow.

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Electronic Supplementary Material

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ESM Table 1

Distribution of BOX-PCR genotypes throughout sampling sites (PDF 24 kb)

ESM Table 2

Population-wide recA and phzF nucleotide sequence distance (a) and diversity calculations (b; PDF 25 kb)

ESM Table 3

Sixty-two (as determined by principle components analysis) Biolog substrates used for cluster analysis (PDF 261 kb)

ESM Table 4

Utilization data for 17 common wheat rhizosphere exudates extracted from Biolog GN2 microplate dataset (PDF 142 kb)

ESM Table 5

Production of phenazine-1-carboxylic acid and biosurfactants by 31 Phz+ genotypes (PDF 20 kb)

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Parejko, J.A., Mavrodi, D.V., Mavrodi, O.V. et al. Population Structure and Diversity of Phenazine-1-Carboxylic Acid Producing Fluorescent Pseudomonas spp. from Dryland Cereal Fields of Central Washington State (USA). Microb Ecol 64, 226–241 (2012). https://doi.org/10.1007/s00248-012-0015-0

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