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Applied and Environmental Microbiology, July 2002, p. 3328-3338, Vol. 68, No. 7
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.7.3328-3338.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Plant-Dependent Genotypic and Phenotypic Diversity of Antagonistic Rhizobacteria Isolated from Different Verticillium Host Plants

Gabriele Berg,1* Nicolle Roskot,1 Anette Steidle,2 Leo Eberl,2 Angela Zock,3 and Kornelia Smalla3

Department of Microbiology, University of Rostock, D-18055 Rostock,1 Department of Microbiology, Technical University of Munich, D-85350 Freising,2 Federal Biological Research Center for Agriculture and Forestry, D-38104 Braunschweig, Germany3

Received 10 December 2001/ Accepted 11 April 2002

To study the effect of plant species on the abundance and diversity of bacterial antagonists, the abundance, the phenotypic diversity, and the genotypic diversity of rhizobacteria isolated from potato, oilseed rape, and strawberry and from bulk soil which showed antagonistic activity towards the soilborne pathogen Verticillium dahliae Kleb. were analyzed. Rhizosphere and soil samples were taken five times over two growing seasons in 1998 and 1999 from a randomized field trial. Bacterial isolates were obtained after plating on R2A (Difco, Detroit, Mich.) or enrichment in microtiter plates containing high-molecular-weight substrates followed by plating on R2A. A total of 5,854 bacteria isolated from the rhizosphere of strawberry, potato, or oilseed rape or bulk soil from fallow were screened by dual testing for in vitro antagonism towards Verticillium. The proportion of isolates with antagonistic activity was highest for strawberry rhizosphere (9.5%), followed by oilseed rape (6.3%), potato (3.7%), and soil (3.3%). The 331 Verticillium antagonists were identified by their fatty acid methyl ester profiles. They were characterized by testing their in vitro antagonism against other pathogenic fungi; their glucanolytic, chitinolytic, and proteolytic activities; and their BOX-PCR fingerprints. The abundance and composition of Verticillium antagonists was plant species dependent. A rather high proportion of antagonists from the strawberry rhizosphere was identified as Pseudomonas putida B (69%), while antagonists belonging to the Enterobacteriaceae (Serratia spp., Pantoea agglomerans) were mainly isolated from the rhizosphere of oilseed rape. For P. putida A and B plant-specific genotypes were observed, suggesting that these bacteria were specifically enriched in each rhizosphere.

* Corresponding author. Mailing address: Universität Rostock, Fachbereich Biowissenschaften, Institut für Molekulare Physiologie und Biotechnologie, Mikrobiologie, D-18051 Rostock, Germany. Phone: 49 381 4986154. Fax: 49 381 4986152. E-mail: gabriele.berg{at}biologie.uni-rostock.de.

Applied and Environmental Microbiology, July 2002, p. 3328-3338, Vol. 68, No. 7
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.7.3328-3338.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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