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 Barbieri, E.
Right arrow Articles by Stocchi, V.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Barbieri, E.
Right arrow Articles by Stocchi, V.
Agricola
Right arrow Articles by Barbieri, E.
Right arrow Articles by Stocchi, V.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2000, p. 5035-5042, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Phylogenetic Characterization and In Situ Detection of a Cytophaga-Flexibacter-Bacteroides Phylogroup Bacterium in Tuber borchii Vittad. Ectomycorrhizal Mycelium

Elena Barbieri,1 Lucia Potenza,1 Ismaela Rossi,1 Davide Sisti,2 Giovanna Giomaro,2 Simona Rossetti,3 Claudia Beimfohr,4 and Vilberto Stocchi1,*

"Giorgio Fornaini" Institute of Biochemistry1 and Institute of Botany,2 University of Urbino, 61029 Urbino, and Water Research Institute, CNR, 00198 Rome,3 Italy, and Vermicon AG, 80992 Munich, Germany4

Received 29 September 1999/Accepted 2 August 2000

Mycorrhizal ascomycetous fungi are obligate ectosymbionts that colonize the roots of gymnosperms and angiosperms. In this paper we describe a straightforward approach in which a combination of morphological and molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria associated with the ascomycetous ectomycorrhizal fungus Tuber borchii Vittad. Universal eubacterial primers specific for the 5' and 3' ends of the 16S rRNA gene (16S rDNA) were used for PCR amplification, direct sequencing, and phylogenetic analyses. The 16S rDNA was amplified directly from four pure cultures of T. borchii Vittad. mycelium. A nearly full-length sequence of the gene coding for the prokaryotic small-subunit rRNA was obtained from each T. borchii mycelium studied. The 16S rDNA sequences were almost identical (98 to 99% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belonging to the Cytophaga-Flexibacter-Bacteroides (CFB) phylogroup which had not been described previously. In situ detection of the CFB bacterium in the hyphal tissue of the fungus T. borchii was carried out by using 16S rRNA-targeted oligonucleotide probes for the eubacterial domain and the Cytophaga-Flexibacter phylum, as well as a probe specifically designed for the detection of this mycelium-associated bacterium. Fluorescent in situ hybridization showed that all three of the probes used bound to the mycelium tissue. This study provides the first direct visual evidence of a not-yet-cultured CFB bacterium associated with a mycorrhizal fungus of the genus Tuber.

* Corresponding author. Mailing address: "Giorgio Fornaini" Institute of Biochemistry, University of Urbino, Via Saffi, 2, 61029 Urbino (PU), Italy. Phone: 39 0722 305262. Fax: 39 0722 320188. E-mail: vstocchi{at}uniurb.it.

Applied and Environmental Microbiology, November 2000, p. 5035-5042, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Bertaux, J., Schmid, M., Prevost-Boure, N. C., Churin, J. L., Hartmann, A., Garbaye, J., Frey-Klett, P. (2003). In Situ Identification of Intracellular Bacteria Related to Paenibacillus spp. in the Mycelium of the Ectomycorrhizal Fungus Laccaria bicolor S238N. Appl. Environ. Microbiol. 69: 4243-4248 [Abstract] [Full Text]  
  • Barbieri, E., Riccioni, G., Pisano, A., Sisti, D., Zeppa, S., Agostini, D., Stocchi, V. (2002). Competitive PCR for Quantitation of a Cytophaga-Flexibacter-Bacteroides Phylum Bacterium Associated with the Tuber borchii Vittad. Mycelium. Appl. Environ. Microbiol. 68: 6421-6424 [Abstract] [Full Text]  
  • Ashelford, K. E., Weightman, A. J., Fry, J. C. (2002). PRIMROSE: a computer program for generating and estimating the phylogenetic range of 16S rRNA oligonucleotide probes and primers in conjunction with the RDP-II database. Nucleic Acids Res 30: 3481-3489 [Abstract] [Full Text]  
  • Zchori-Fein, E., Gottlieb, Y., Kelly, S. E., Brown, J. K., Wilson, J. M., Karr, T. L., Hunter, M. S. (2001). A newly discovered bacterium associated with parthenogenesis and a change in host selection behavior in parasitoid wasps. Proc. Natl. Acad. Sci. USA 10.1073/pnas.221467498v1 [Abstract] [Full Text]  
  • Zchori-Fein, E., Gottlieb, Y., Kelly, S. E., Brown, J. K., Wilson, J. M., Karr, T. L., Hunter, M. S. (2001). A newly discovered bacterium associated with parthenogenesis and a change in host selection behavior in parasitoid wasps. Proc. Natl. Acad. Sci. USA 98: 12555-12560 [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»