Published: Vol 5, Iss 16, Aug 20, 2015 DOI: 10.21769/BioProtoc.1569 Views: 25872
Reviewed by: Zhaohui LiuYurong XieAnonymous reviewer(s)
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Abstract
Rhizosphere bacterial communities have become a major focal point of research in recent years, especially regarding how they affect plants and vice versa (Philippot et al., 2013). Changes in microbial density and diversity within the rhizosphere occur in a spatial temporal manner. The soil zone closest to the plant roots has the most density and diversity of microbes (Clark, 1940). The lack of methods to consistently isolate rhizosphere samples in a spatially defined manner is a major bottleneck in rhizosphere microbiology. We hypothesized that microbes with increasing affinities to and distance from the plant root can be isolated using increasing strengths of physical disruption. Sonication is an excellent choice due to the ability to gently remove rhizosphere soil and bacterial biofilms without damaging plant roots (Doi T et al., 2007; Bulgarelli et al., 2012; Lundberg et al., 2012). In addition, simply increasing the time of sonication can increase the amount of physical force. We used such an approach to consistently isolate microbial communities with different affinities to the soybean roots (White et al., 2014). This article describes the use of successive sonication to isolate distal, middle, and proximal soil from the rhizosphere of soybean roots.
Materials and Reagents
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Limitations of the method
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Acknowledgments
This protocol was established in a previously published study (White et al., 2014). Funding for this research was provided by the South Dakota Agricultural Experiment Station and the South Dakota Soybean Research and Promotion Council. We would also like to thank R. Gelderman (SDSU) for providing soil samples, M. Hildreth (SDSU) for providing the sonicator used for this research, and Al Miron for providing the soybean plant depicted in Figure 1.
References
Article Information
Copyright
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
White, L. J., Brözel, V. S. and Subramanian, S. (2015). Isolation of Rhizosphere Bacterial Communities from Soil. Bio-protocol 5(16): e1569. DOI: 10.21769/BioProtoc.1569.
Category
Microbiology > Microbial cell biology > Cell isolation and culture
Microbiology > Microbe-host interactions > Bacterium
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