Techniques to Study Common Root Responses to Beneficial Microbes and Iron Deficiency

Shu Hua Hsu; Max J.J. Stassen; Corné M.J. Pieterse; Ioannis A. Stringlis

doi:10.1007/978-1-0716-3183-6_5

Techniques to Study Common Root Responses to Beneficial Microbes and Iron Deficiency

Shu Hua Hsu, Max J.J. Stassen, Corné M.J. Pieterse, Ioannis A. Stringlis^*

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

Abstract

Iron (Fe) plays a central role in the vital processes of a plant. The Fe status of a plant influences growth and immunity, but it also dictates interactions of roots with soil microbiota through the production of Fe mobilizing, antimicrobial fluorescent phenolic compounds called coumarins. To adapt to low Fe availability in the soil, plants deploy an efficient Fe deficiency response. Interestingly, this Fe deficiency response is hijacked by root-colonizing microbes in the root microbiome to establish a mutually beneficial relationship. In this chapter, we describe how we cultivate plants and microbes to study the interaction between plants, beneficial rhizobacteria, and the plant’s Fe deficiency response. We describe (a) how we study activity and localization of these responses by assessing gene-specific promoter activities using GUS assays, (b) how we visualize root-secreted coumarins in response to Fe deficiency and colonization by beneficial rhizobacteria, and (c) how we prepare our samples for metabolite extraction and reverse-transcriptase quantitative PCR to analyze the expression of marker genes.

Original language	English
Title of host publication	Methods in Molecular Biology
Subtitle of host publication	Plant Iron Homeostasis: Methods and Protocols
Publisher	Humana Press
Pages	47-62
Number of pages	16
Volume	2665
DOIs	https://doi.org/10.1007/978-1-0716-3183-6_5
Publication status	Published - 2023

Publication series

Name	Methods in Molecular Biology
Volume	2665
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Funding

The authors would like to thank Hans van Pelt for the high quality pictures of the experiments and the members of the Plant-Microbe Interactions group who had developed some of the preceding protocols. This work is supported by NWO Gravitation Grant no. 662 024.004.014.

Funders	Funder number
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	662 024.004.014

Keywords

Coumarins
Induced systemic resistance
Iron deficiency
Plant-microbes interaction
Pseudomonas simiae WCS417r

Access to Document

10.1007/978-1-0716-3183-6_5

MethodsMolBiol-Hsu-2023Final published version, 14.3 MBLicence: Taverne

Cite this

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abstract = "Iron (Fe) plays a central role in the vital processes of a plant. The Fe status of a plant influences growth and immunity, but it also dictates interactions of roots with soil microbiota through the production of Fe mobilizing, antimicrobial fluorescent phenolic compounds called coumarins. To adapt to low Fe availability in the soil, plants deploy an efficient Fe deficiency response. Interestingly, this Fe deficiency response is hijacked by root-colonizing microbes in the root microbiome to establish a mutually beneficial relationship. In this chapter, we describe how we cultivate plants and microbes to study the interaction between plants, beneficial rhizobacteria, and the plant{\textquoteright}s Fe deficiency response. We describe (a) how we study activity and localization of these responses by assessing gene-specific promoter activities using GUS assays, (b) how we visualize root-secreted coumarins in response to Fe deficiency and colonization by beneficial rhizobacteria, and (c) how we prepare our samples for metabolite extraction and reverse-transcriptase quantitative PCR to analyze the expression of marker genes.",

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Techniques to Study Common Root Responses to Beneficial Microbes and Iron Deficiency. / Hsu, Shu Hua ; Stassen, Max J.J.; Pieterse, Corné M.J. et al.
Methods in Molecular Biology: Plant Iron Homeostasis: Methods and Protocols. Vol. 2665 Humana Press, 2023. p. 47-62 (Methods in Molecular Biology; Vol. 2665).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

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Techniques to Study Common Root Responses to Beneficial Microbes and Iron Deficiency

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