This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wullings, B. A.
Right arrow Articles by Akkermans, A. D. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wullings, B. A.
Right arrow Articles by Akkermans, A. D. L.
Agricola
Right arrow Articles by Wullings, B. A.
Right arrow Articles by Akkermans, A. D. L.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 1998, p. 4546-4554, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Detection of Ralstonia solanacearum, Which Causes Brown Rot of Potato, by Fluorescent In Situ Hybridization with 23S rRNA-Targeted Probes

B. A. Wullings,1,dagger A. R. Van Beuningen,2 J. D. Janse,2 and A. D. L. Akkermans1,*

Laboratory of Microbiology, Department of Biomolecular Sciences, Wageningen Agricultural University,1 and Department of Bacteriology, Plant Protection Service,2 Wageningen, The Netherlands

Received 23 February 1998/Accepted 18 July 1998

During the past few years, Ralstonia (Pseudomonas) solanacearum race 3, biovar 2, was repeatedly found in potatoes in Western Europe. To detect this bacterium in potato tissue samples, we developed a method based on fluorescent in situ hybridization (FISH). The nearly complete genes encoding 23S rRNA of five R. solanacearum strains and one Ralstonia pickettii strain were PCR amplified, sequenced, and analyzed by sequence alignment. This resulted in the construction of an unrooted tree and supported previous conclusions based on 16S rRNA sequence comparison in which R. solanacearum strains are subdivided into two clusters. Based on the alignments, two specific probes, RSOLA and RSOLB, were designed for R. solanacearum and the closely related Ralstonia syzygii and blood disease bacterium. The specificity of the probes was demonstrated by dot blot hybridization with RNA extracted from 88 bacterial strains. Probe RSOLB was successfully applied in FISH detection with pure cultures and potato tissue samples, showing a strong fluorescent signal. Unexpectedly, probe RSOLA gave a less intense signal with target cells. Potato samples are currently screened by indirect immunofluorescence (IIF). By simultaneously applying IIF and the developed specific FISH, two independent targets for identification of R. solanacearum are combined, resulting in a rapid (1-day), accurate identification of the undesired pathogen. The significance of the method was validated by detecting the pathogen in soil and water samples and root tissue of the weed host Solanum dulcamara (bittersweet) in contaminated areas.

* Corresponding author. Mailing address: Laboratory of Microbiology, Wageningen Agricultural University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands. Phone: 31 317 483486. Fax: 31 317 483829. E-mail: Antoon.Akkermans{at}algemeen.micr.wau.nl.

dagger Present address: KIWA N.V., Research and Consultancy, Nieuwegein, The Netherlands.

Applied and Environmental Microbiology, November 1998, p. 4546-4554, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Girlich, D., Naas, T., Nordmann, P. (2004). OXA-60, a Chromosomal, Inducible, and Imipenem-Hydrolyzing Class D {beta}-Lactamase from Ralstonia pickettii. Antimicrob. Agents Chemother. 48: 4217-4225 [Abstract] [Full Text]  
  • Bianciotto, V., Genre, A., Jargeat, P., Lumini, E., Becard, G., Bonfante, P. (2004). Vertical Transmission of Endobacteria in the Arbuscular Mycorrhizal Fungus Gigaspora margarita through Generation of Vegetative Spores. Appl. Environ. Microbiol. 70: 3600-3608 [Abstract] [Full Text]  
  • Felske, A., Wolterink, A., Van Lis, R., De Vos, W. M., Akkermans, A. D. L. (2000). Response of a Soil Bacterial Community to Grassland Succession as Monitored by 16S rRNA Levels of the Predominant Ribotypes. Appl. Environ. Microbiol. 66: 3998-4003 [Abstract] [Full Text]  
  • Weller, S. A., Elphinstone, J. G., Smith, N. C., Boonham, N., Stead, D. E. (2000). Detection of Ralstonia solanacearum Strains with a Quantitative, Multiplex, Real-Time, Fluorogenic PCR (TaqMan) Assay. Appl. Environ. Microbiol. 66: 2853-2858 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»