In vivo gibberellin gradients visualized in rapidly elongating tissues

Abstract

The phytohormone gibberellin (GA) is a key regulator of plant growth and development. Although the upstream regulation and downstream responses to GA vary across cells and tissues, developmental stages and environmental conditions, the spatiotemporal distribution of GA in vivo remains unclear. Using a combinatorial screen in yeast, we engineered an optogenetic biosensor, GIBBERELLIN PERCEPTION SENSOR 1 (GPS1), that senses nanomolar levels of bioactive GAs. Arabidopsis thaliana plants expressing a nuclear localized GPS1 report on GAs at the cellular level. GA gradients were correlated with gradients of cell length in rapidly elongating roots and dark-grown hypocotyls. In roots, accumulation of exogenously applied GA also correlated with cell length, intimating that a root GA gradient can be established independently of GA biosynthesis. In hypocotyls, GA levels were reduced in a phytochrome interacting factor (pif) quadruple mutant in the dark and increased in a phytochrome double mutant in the light, indicating that PIFs elevate GA in the dark and that phytochrome inhibition of PIFs could lower GA in the light. As GA signalling directs hypocotyl elongation largely through promoting PIF activity, PIF promotion of GA accumulation represents a positive feedback loop within the molecular framework driving rapid hypocotyl growth.

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Fig. 1: Engineering and GA response of GPS1 and GPS1-NR.
Fig. 2: Fluorescence emission ratio response of purified GPS1 to GAs.
Fig. 3: Emission ratios of nlsGPS1 in root tips.
Fig. 4: Emission ratios of nlsGPS1 in roots before and after treatment with GAs.
Fig. 5: Emission ratios of nlsGPS1 in light-grown hypocotyls.
Fig. 6: Relationship between cell size and nlsGPS1 emission ratio.

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Acknowledgements

We gratefully acknowledge S. Manoj-Kumar for technical assistance and B. DeMeo for development of XT Mean Intensity Ratio. We thank the Gatsby Charitable Foundation for funding support of A.R., A.W. and A.M.J.

Author information

A.R., A.W., V.L. and A.M.J. ran the experiments. A.R., A.W., V.L. and A.M.J. analysed the results. A.R. and A.M.J. wrote the manuscript, and W.B.F. and A.M.J. designed and supervised the project.

Correspondence to Wolf B. Frommer or Alexander M. Jones.

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Supplementary Information

Supplementary Figures 1–11, Supplementary Table 1.

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Supplementary Video 1

Time-course of nlsGPS1 emission ratios of an Arabidopsis root tip growing in the RootChip16. Time between frames was 5 minutes. Perfusion environment was as follows: 15 minutes mock solution, 20 minutes 100 nM GA4, 120 minutes mock solution, 20 minutes 100 nM GA4, 80 minutes mock solution.Time-course of nlsGPS1 emission ratios of an root tip growing in the RootChip16. Time between frames was 5 minutes. Perfusion environment was as follows: 15 minutes mock solution, 20 minutes 100 nM GA, 120 minutes mock solution, 20 minutes 100 nM GA, 80 minutes mock solution.

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Rizza, A., Walia, A., Lanquar, V. et al. In vivo gibberellin gradients visualized in rapidly elongating tissues. Nature Plants 3, 803–813 (2017). https://doi.org/10.1038/s41477-017-0021-9

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