期刊
MICROGRAVITY SCIENCE AND TECHNOLOGY
卷 28, 期 1, 页码 59-66出版社
SPRINGER
DOI: 10.1007/s12217-015-9478-1
关键词
Arabidopsis thaliana; Clinostat; Gene expression; Gravity; Auxin; Signal transduction
资金
- Excellence Initiative of the German Federal and State Governments [EXC 294]
- European Space Agency
- German Aerospace Center, Space Administration on behalf of the Bundesministerium fur Wirtschaft und Technologie (BMWi)
- National Natural Science Foundation of China [31570291]
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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