Quantifying Phytohormones in Vivo with FRET Biosensors and the FRETENATOR Analysis Toolset

James H Rowe; Annalisa Rizza; Alexander M Jones

doi:10.1007/978-1-0716-2297-1_17

Quantifying Phytohormones in Vivo with FRET Biosensors and the FRETENATOR Analysis Toolset

Methods Mol Biol. 2022:2494:239-253. doi: 10.1007/978-1-0716-2297-1_17.

Authors

James H Rowe¹, Annalisa Rizza¹, Alexander M Jones²

Affiliations

¹ Sainsbury Laboratory, University of Cambridge, Cambridge, UK.
² Sainsbury Laboratory, University of Cambridge, Cambridge, UK. alexander.jones@slcu.cam.ac.uk.

PMID: 35467212
DOI: 10.1007/978-1-0716-2297-1_17

Abstract

The ABACUS1-2 μ (ABscisic Acid Concentration and Uptake Sensor 1-2 μ) and GPS1 (Gibberellin Perception Sensor 1) are direct Förster resonance energy transfer (FRET) biosensors that can be used to measure hormone levels in planta. We provide detailed protocols to image FRET biosensors under exogenously applied hormones in roots, either as a single time point or for treatment time courses before and after hormone application. A new, free, open-source analysis toolset for Fiji is introduced and used to get full 3D segmentation of images of nuclear localized FRET biosensors and calculate emission ratios on a per nucleus basis allowing in-depth analysis of biosensor data.

Keywords: ABA; Abscisic acid; Biosensors; FRET; GA; Gibberellin; Image analysis; Microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biosensing Techniques* / methods
Fluorescence Resonance Energy Transfer* / methods
Gibberellins
Hormones
Plant Growth Regulators

Substances

Gibberellins
Hormones
Plant Growth Regulators

Grants and funding

BB/P018572/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom