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Microcosm Approaches to Investigate Multitrophic Interactions between Microbial Communities in the Rhizosphere of Plants

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Methods in Rhizosphere Biology Research

Part of the book series: Rhizosphere Biology ((RHBIO))

Abstract

Plant roots are constantly forced to interact with a multitude of root colonising and free-living microorganisms in the rhizosphere. Rhizosphere microbiology, however, is traditionally split into several sub-disciplines, focusing e.g. on mycorrhiza, other endophytic fungi, bacteria, ‘plant growth promoting bacteria’, protists, ‘root fungal pathogens’, root nematodes, etc.. Accordingly, many plant-microbe studies exist on single organism groups. Combining different microbial groups in an experimental study is technically challenging because of the complexity of the set up, timing of inoculation and risk of contaminations. This chapter is intended as a step-by-step manual for the set-up of multitrophic experiments with microorganisms and plants. Technical challenges will be discussed at each step. I will first discuss different methods of soil sterilization. Then I will present different isolation, cultivation and inoculation techniques that we developed or adapted for multitrophic interaction studies. Finally, I present different microcosm systems designed to investigate specific interactions in plant-microbe studies.

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Acknowlegements

This work was supported by the Cluster of Excellence on Plant Sciences (CEPLAS), and the priority program of the German Science Foundation DFG-SPP2089 “Rhizosphere Spatiotemporal Organisation”. We are particularly grateful for the continuous contributions and support of Leo Leson and the whole workshop team of the Biocenter, University of Cologne.

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Authors and Affiliations

  1. Terrestrial Ecology, Institute of Zoology, University of Cologne, Cologne, Germany

    Michael Bonkowski

  2. Cluster of Excellence on Plant Sciences (CEPLAS), University of Cologne, Cologne, Germany

    Michael Bonkowski

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  1. Michael Bonkowski
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Corresponding author

Correspondence to Michael Bonkowski .

Editor information

Editors and Affiliations

  1. Department of Biology, University of Fribourg, Fribourg, Switzerland

    Didier Reinhardt

  2. Biological Sciences, CBSH, G.B.Pant University of Agriculture & Technology, Pantnagar, Uttarakhand, India

    Anil K. Sharma

Appendix

Appendix

  1. 1.

    Neff’s modified Amoeba Salinae (NMAS):

  • Stock 1: 12 g NaCl, 0.4 g MgSO4 * 7H2O, 0.6 g CaCl2 * 6 H2O, fill up to 1000 ml with H2Odest.

  • Stock 2: 14.2 g Na2HPO4, 13.6 g KH2PO4, fill up to 1000 ml with H2Odest.

  • Mix 5 ml of stock 1 and 5 ml of stock 2 with 1000 ml of H2Odest and autoclave for 20 min at 120 °C.

  1. 2.

    NB-NMAS (Nutrient Broth – Neff’s Modified Amoebae Salinae):

  • Add 0.8 g nutrient broth (Oxoid, UK) to 1000 ml NMAS. Autoclave for 20 min at 120 °C.

  1. 3.

    Prescott’s James’s solution (PJ):

    Stock solution 1

    Add 0.433 g CaCl2·2H2O and 0.162 g KCl to 100 ml H2O

    Stock solution 2

    Add 0.512 g K2HPO4 to 100 ml H2O

    Stock solution 3

    Add 0.280 g MgSO4 to100 ml H2O

One ml of the Stock solutions 1, 2 and 3 each is filled up to 1000 ml with H2Odest. Autoclave for 20 min at 120 °C.

  1. 4.

    Wheat Grass Medium (WG):

  • 0,15 g wheat grass powder are three times boiled up with 200 ml H2Odest, filtered and added to 1000 ml PJ solution. Autoclave for 20 min at 120 °C.

  1. 5.

    Gamborg medium (for plants):

  • Add 3.2 g of Gamborg’s B-5 basal medium with minimal organics to 1000 ml H2Odest. Autoclave for 20 min at 120 °C.

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Bonkowski, M. (2019). Microcosm Approaches to Investigate Multitrophic Interactions between Microbial Communities in the Rhizosphere of Plants. In: Reinhardt, D., Sharma, A. (eds) Methods in Rhizosphere Biology Research. Rhizosphere Biology. Springer, Singapore. https://doi.org/10.1007/978-981-13-5767-1_14

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