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- Title
In vivo monitoring of plant small GTPase activation using a Förster resonance energy transfer biosensor.
- Authors
Wong, Hann Ling; Akamatsu, Akira; Wang, Qiong; Higuchi, Masayuki; Matsuda, Tomonori; Okuda, Jun; Kosami, Ken-ichi; Inada, Noriko; Kawasaki, Tsutomu; Kaneko-Kawano, Takako; Nagawa, Shingo; Tan, Li; Kawano, Yoji; Shimamoto, Ko
- Abstract
Background: Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patterning and hormone responses. Thus, to monitor their activation status within plant cells is believed to be the key step in understanding their roles. Results: We have established a plant version of a Förster resonance energy transfer (FRET) probe called Ras and interacting protein chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 during various defence responses in cells. Here, we describe the protocol for visualizing spatiotemporal activity of plant Rac/ROP GTPase in living plant cells, transfection of rice protoplasts with <italic>Raichu</italic>-<italic>OsRac1</italic> and acquisition of FRET images. Conclusions: Our protocol should be adaptable for monitoring activation for other plant small GTPases and protein–protein interactions for other FRET sensors in various plant cells.
- Subjects
GUANOSINE triphosphatase; MOLECULAR switches; BIOSENSORS; IN vivo studies; DISEASE resistance of plants
- Publication
Plant Methods, 2018, Vol 14, Issue 1, pN.PAG
- ISSN
1746-4811
- Publication type
Article
- DOI
10.1186/s13007-018-0325-4