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Applied and Environmental Microbiology, May 2000, p. 1939-1946, Vol. 66, No. 5
0099-2240/00/$04.00+0

Genotypic and Phenotypic Diversity of phlD-Containing Pseudomonas Strains Isolated from the Rhizosphere of Wheat

B. B. McSpadden Gardener,1,* K. L. Schroeder,1 S. E. Kalloger,1 J. M. Raaijmakers,2 L. S. Thomashow,1 and D. M. Weller1

Root Disease and Biological Control Research Unit, USDA Agricultural Research Service, Washington State University, Pullman, Washington,1 and Department of Phytopathology, Wageningen Agricultural University, Wageningen, The Netherlands2

Received 8 October 1999/Accepted 2 March 2000

Production of 2,4-diacetylphloroglucinol (2,4-DAPG) in the rhizosphere by strains of fluorescent Pseudomonas spp. results in the suppression of root diseases caused by certain fungal plant pathogens. In this study, fluorescent Pseudomonas strains containing phlD, which is directly involved in the biosynthesis of 2,4-DAPG, were isolated from the rhizosphere of wheat grown in soils from wheat-growing regions of the United States and The Netherlands. To assess the genotypic and phenotypic diversity present in this collection, 138 isolates were compared to 4 previously described 2,4-DAPG producers. Thirteen distinct genotypes, one of which represented over 30% of the isolates, were differentiated by whole-cell BOX-PCR. Representatives of this group were isolated from eight different soils taken from four different geographic locations. ERIC-PCR gave similar results overall, differentiating 15 distinct genotypes among all of the isolates. In most cases, a single genotype predominated among isolates obtained from each soil. Thirty isolates, representing all of the distinct genotypes and geographic locations, were further characterized. Restriction analysis of amplified 16S rRNA gene sequences revealed only three distinct phylogenetic groups, one of which accounted for 87% of the isolates. Phenotypic analyses based on carbon source utilization profiles revealed that all of the strains utilized 49 substrates and were unable to grow on 12 others. Individually, strains could utilize about two-thirds of the 95 substrates present in Biolog SF-N plates. Multivariate analyses of utilization profiles revealed phenotypic groupings consistent with those defined by the genotypic analyses.

* Corresponding author. Mailing address: Root Disease and Biological Control Research Unit, USDA Agricultural Research Service, Pullman, WA 99164-6430. Phone: (509) 335-1116. Fax: (509) 335-7674. E-mail: mcspadde{at}mail.wsu.edu.

Applied and Environmental Microbiology, May 2000, p. 1939-1946, Vol. 66, No. 5
0099-2240/00/$04.00+0


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