Abstract
Plant growth relates to gravity, ensuring that roots grow downwards into the soil and shoots expand aerially. The phytohormone auxin mediates tropistic growth responses, such as root gravitropism. Gravity perception in the very tip of the roots triggers carrier-dependent, asymmetric redistribution of auxin, leading to differential auxin responses and growth regulation at the upper and lower root flanks. This cellular, asymmetry-breaking event will eventually lead to root bending towards the gravity vector. Here, we show how to investigate auxin signaling and auxin carrier dynamics during root gravitropic response, using a chambered cover glass in combination with a confocal live cell imaging approach. To exemplify this method, we used established lines expressing transcriptional and translational green fluorescent protein (GFP) fusions to the auxin responsive promoter element DR5rev and the prominent auxin carrier PIN-FORMED2 (PIN2), respectively. Transgenic seedlings were placed and grown in the chambered cover glasses, enabling defined gravitropic stimulations prior to imaging. This method is optimal for inverted microscopes and significantly reduces stressful manipulations during specimen preparation.
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Acknowledgements
We are grateful to B. Scheres and J. Friml for providing the published fluorescent marker lines; the BOKU-VIBT Imaging Center for access and C. Löfke for expertise; D. Whittaker for help in preparing the manuscript. This work was supported by the Vienna Science and Technology Fund (WWTF) (to J.K.-V.), Austrian Science Fund (FWF) (Projects: P26568-B16 and P26591-B16 to J.K.-V.), and European Molecular Biology Organization (EMBO) (ALTF 795-2012, personal postdoctoral research grant to E.F.).
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Feraru, M.I., Kleine-Vehn, J., Feraru, E. (2015). Auxin Carrier and Signaling Dynamics During Gravitropic Root Growth. In: Blancaflor, E. (eds) Plant Gravitropism. Methods in Molecular Biology, vol 1309. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2697-8_7
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DOI: https://doi.org/10.1007/978-1-4939-2697-8_7
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