Abstract
Ground-based simulators of microgravity such as fast rotating 2-D clinostats are valuable tools to study gravity related processes. We describe here a versatile g-value-adjustable 2-D clinostat that is suitable for plant analysis. To avoid seedling adaptation to 1 g after clinorotation, we designed chambers that allow rapid fixation. A detailed protocol for fixation, RNA isolation and the analysis of selected genes is described. Using this clinostat we show that mRNA levels of LONG HYPOCOTYL 5 (HY5), MIZU-KUSSEI 1 (MIZ1) and microRNA MIR163 are down-regulated in 5-day-old Arabidopsis thaliana roots after 3 min and 6 min of clinorotation using a maximal reduced g-force of 0.02 g, hence demonstrating that this 2-D clinostat enables the characterization of early transcriptomic events during root response to microgravity. We further show that this 2-D clinostat is able to compensate the action of gravitational force as both gravitropic-dependent statolith sedimentation and subsequent auxin redistribution (monitoring D R5 r e v ::G F P reporter) are abolished when plants are clinorotated. Our results demonstrate that 2-D clinostats equipped with interchangeable growth chambers and tunable rotation velocity are suitable for studying how plants perceive and respond to simulated microgravity.
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Acknowledgments
This work was supported by the Excellence Initiative of the German Federal and State Governments (EXC 294), the European Space Agency, and the German Aerospace Center, Space Administration on behalf of the Bundesministerium für Wirtschaft und Technologie (BMWi) and National Natural Science Foundation of China (31570291). We thank all members of our team for their help and support.
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Wang, H., Li, X., Krause, L. et al. 2-D Clinostat for Simulated Microgravity Experiments with Arabidopsis Seedlings. Microgravity Sci. Technol. 28, 59–66 (2016). https://doi.org/10.1007/s12217-015-9478-1
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DOI: https://doi.org/10.1007/s12217-015-9478-1