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

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Mar 1995, 1013-1019, Vol 61, No. 3
Copyright © 1995, American Society for Microbiology

In Situ Localization of Azospirillum brasilense in the Rhizosphere of Wheat with Fluorescently Labeled, rRNA-Targeted Oligonucleotide Probes and Scanning Confocal Laser Microscopy

B Assmus, P Hutzler, G Kirchhof, R Amann, JR Lawrence and A Hartmann
Institut fur Bodenokologie and Institut fur Pathologie, biomedizinische Bildanalyse, GSF-Forschungszentrum fur Umwelt und Gesundheit, D-85764 Oberschleissheim, and Lehrstuhl fur Mikrobiologie, Technische Universitat Munchen, D-80290 Munich, Germany, and National Hydrology Research Institute, Environment Canada, Saskatoon, Saskatchewan, Canada S7N 3H5

The colonization of wheat roots by Azospirillum brasilense was used as a model system to evaluate the utility of whole-cell hybridization with fluorescently labeled, rRNA-targeted oligonucleotide probes for the in situ monitoring of rhizosphere microbial communities. Root samples of agar- or soil-grown 10- and 30-day-old wheat seedlings inoculated with different strains of A. brasilense were hybridized with a species-specific probe for A. brasilense, a probe hybridizing to alpha subclass proteobacteria, and a probe specific for the domain Bacteria to identify and localize the target bacteria. After hybridization, about 10 to 25% of the rhizosphere bacteria as visualized with 4(prm1),6-diamidino-2-phenylindole (DAPI) gave sufficient fluorescence signals to be detected with rRNA-targeted probes. Scanning confocal laser microscopy was used to overcome disturbing effects arising from autofluorescence of the object or narrow depth of focus in thick specimens. This technique also allowed high-resolution analysis of the spatial distribution of bacteria in the rhizosphere. Occurrence of cells of A. brasilense Sp7 and Wa3 was restricted to the rhizosphere soil, mainly to the root hair zone. C-forms of A. brasilense were demonstrated to be physiologically active forms in the rhizosphere. Strain Sp245 also was found repeatedly at high density in the interior of root hair cells. In general, the combination of fluorescently labeled oligonucleotide probes and scanning confocal laser microscopy provided a very suitable strategy for detailed studies of rhizosphere microbial ecology.

This article has been cited by other articles:

• Rogers, S. W., Moorman, T. B., Ong, S. K. (2007). Fluorescent In Situ Hybridization and Micro-autoradiography Applied to Ecophysiology in Soil. Soil Sci. 71: 620-631 [Abstract] [Full Text]  
• Eller, G., Frenzel, P. (2001). Changes in Activity and Community Structure of Methane-Oxidizing Bacteria over the Growth Period of Rice. Appl. Environ. Microbiol. 67: 2395-2403 [Abstract] [Full Text]  
• Weber, S., Stubner, S., Conrad, R. (2001). Bacterial Populations Colonizing and Degrading Rice Straw in Anoxic Paddy Soil. Appl. Environ. Microbiol. 67: 1318-1327 [Abstract] [Full Text]  
• Normander, B., Prosser, J. I. (2000). Bacterial Origin and Community Composition in the Barley Phytosphere as a Function of Habitat and Presowing Conditions. Appl. Environ. Microbiol. 66: 4372-4377 [Abstract] [Full Text]  
• Pereg Gerk, L., Gilchrist, K., Kennedy, I. R. (2000). Mutants with Enhanced Nitrogenase Activity in Hydroponic Azospirillum brasilense-Wheat Associations. Appl. Environ. Microbiol. 66: 2175-2184 [Abstract] [Full Text]  
• Chelius, M. K., Triplett, E. W. (2000). Immunolocalization of Dinitrogenase Reductase Produced by Klebsiella pneumoniae in Association with Zea mays L.. Appl. Environ. Microbiol. 66: 783-787 [Abstract] [Full Text]  
• Ramos, C., Mølbak, L., Molin, S. (2000). Bacterial Activity in the Rhizosphere Analyzed at the Single-Cell Level by Monitoring Ribosome Contents and Synthesis Rates. Appl. Environ. Microbiol. 66: 801-809 [Abstract] [Full Text]  
• Yang, C.-H., Crowley, D. E. (2000). Rhizosphere Microbial Community Structure in Relation to Root Location and Plant Iron Nutritional Status. Appl. Environ. Microbiol. 66: 345-351 [Abstract] [Full Text]  
• Tombolini, R., van der Gaag, D. J., Gerhardson, B., Jansson, J. K. (1999). Colonization Pattern of the Biocontrol Strain Pseudomonas chlororaphis MA 342 on Barley Seeds Visualized by Using Green Fluorescent Protein. Appl. Environ. Microbiol. 65: 3674-3680 [Abstract] [Full Text]  
• Lee, N., Nielsen, P. H., Andreasen, K. H., Juretschko, S., Nielsen, J. L., Schleifer, K.-H., Wagner, M. (1999). Combination of Fluorescent In Situ Hybridization and Microautoradiography---a New Tool for Structure-Function Analyses in Microbial Ecology. Appl. Environ. Microbiol. 65: 1289-1297 [Abstract] [Full Text]