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Applied and Environmental Microbiology, June 2009, p. 3396-3406, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.00491-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Endophytic Colonization of Potato (Solanum tuberosum L.) by a Novel Competent Bacterial Endophyte, Pseudomonas putida Strain P9, and Its Effect on Associated Bacterial Communities{triangledown}

Fernando Dini Andreote,1,2 Welington L. de Araújo,2 João L. de Azevedo,2 Jan Dirk van Elsas,3 Ulisses Nunes da Rocha,1 and Leonard S. van Overbeek1*

Plant Research International B.V., Wageningen, The Netherlands,1 Department of Genetics, Escola Superior de Agricultura "Luiz de Queiroz," University of São Paulo, Piracicaba, Brazil,2 Department of Microbial Ecology, Centre for Ecological and Evolutionary Studies, Groningen University, Groningen, The Netherlands3

Received 27 February 2009/ Accepted 24 March 2009

Pseudomonas putida strain P9 is a novel competent endophyte from potato. P9 causes cultivar-dependent suppression of Phytophthora infestans. Colonization of the rhizoplane and endosphere of potato plants by P9 and its rifampin-resistant derivative P9R was studied. The purposes of this work were to follow the fate of P9 inside growing potato plants and to establish its effect on associated microbial communities. The effects of P9 and P9R inoculation were studied in two separate experiments. The roots of transplants of three different cultivars of potato were dipped in suspensions of P9 or P9R cells, and the plants were planted in soil. The fate of both strains was followed by examining colony growth and by performing PCR-denaturing gradient gel electrophoresis (PCR-DGGE). Colonies of both strains were recovered from rhizoplane and endosphere samples of all three cultivars at two growth stages. A conspicuous band, representing P9 and P9R, was found in all Pseudomonas PCR-DGGE fingerprints for treated plants. The numbers of P9R CFU and the P9R-specific band intensities for the different replicate samples were positively correlated, as determined by linear regression analysis. The effects of plant growth stage, genotype, and the presence of P9R on associated microbial communities were examined by multivariate and unweighted-pair group method with arithmetic mean cluster analyses of PCR-DGGE fingerprints. The presence of strain P9R had an effect on bacterial groups identified as Pseudomonas azotoformans, Pseudomonas veronii, and Pseudomonas syringae. In conclusion, strain P9 is an avid colonizer of potato plants, competing with microbial populations indigenous to the potato phytosphere. Bacterization with a biocontrol agent has an important and previously unexplored effect on plant-associated communities.

* Corresponding author. Mailing address: Plant Research International, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands. Phone: 31(0)317 480606. Fax: 31(0)317 418094. E-mail: l.s.vanoverbeek{at}wur.nl

{triangledown} Published ahead of print on 27 March 2009.

Applied and Environmental Microbiology, June 2009, p. 3396-3406, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.00491-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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