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  • A gnotobiotic growth assay for Arabidopsis root microbiota reconstitution under iron limitation.

A gnotobiotic growth assay for Arabidopsis root microbiota reconstitution under iron limitation.

STAR protocols (2020-12-31)
Christopher J Harbort, Masayoshi Hashimoto, Haruhiko Inoue, Paul Schulze-Lefert
ABSTRACT

We present a gnotobiotic system for microbiota reconstitution on Arabidopsis thaliana under contrasting iron availability. This system induces iron starvation in plants by providing an unavailable form, mimicking conditions in alkaline soils. Inoculation of taxonomically diverse bacteria reconstitutes plants with a synthetic microbiota, allowing observation of nutrient-dependent interactions with commensals. Experimental optimization, including media composition and preparation of seedlings and bacteria, is discussed. This system provides a framework that can be adapted to study plant-microbiota interactions in further nutritional contexts. For complete details on the use and execution of this protocol, please refer to Harbort et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Boric acid, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
Sigma-Aldrich
Ammonium nitrate, ≥99.0%
Sigma-Aldrich
Zinc chloride, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid ferric sodium salt, BioReagent, suitable for plant cell culture, crystalline
Sigma-Aldrich
Manganese(II) chloride tetrahydrate, BioReagent, suitable for insect cell culture
Sigma-Aldrich
Copper(II) chloride dihydrate, powder, BioReagent, suitable for cell culture