In vivo monitoring of plant small GTPase activation using a F?rster resonance energy transfer biosensor
In vivo monitoring of plant small GTPase activation using a F?rster resonance energy transfer biosensor
ABSTRACT 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. We have established a plant version of a F?rster 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 rice cells. Here, we describe the protocol for visualizing spatiotemporal activity of plant Rac/ROP GTPase in living plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be widely adaptable for monitoring activation for other plant small GTPases and for other FRET sensors in various plant cells.
Kosami Ken-ichi、Kawasaki Tsutomu、Nagawa Shingo、Shimamoto Ko、Wong Hann Ling、Wang Qiong、Higuchi Masayuki、Inada Noriko、Okuda Jun、Matsuda Tomonori、Akamatsu Akira、Kawano Yoji、Tan Li
Shanghai Center for Plant Stress Biology, Chinese Academy of SciencesLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and Technology||Department of Advanced Bioscience, Kinki UniversityCore Facility of Cell biology, Shanghai Center for Plant Stress BiologyLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and TechnologyLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and Technology||Department of Biological Science, University Tunku Abdul Rahman, Jalan UniversitiShanghai Center for Plant Stress Biology, Chinese Academy of SciencesLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and TechnologyPlant Global Education Project, Nara Institute of Science and Technology||Laboratory of Plant Growth Regulation, Graduate School of Biological Sciences, Nara Institute of Science and TechnologyLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and TechnologyLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and TechnologyLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and Technology||Cell and Developmental Biology, John Innes CentreLaboratory of Plant Molecular Genetics, Graduate School of Biological Sciences, Nara Institute of Science and Technology||Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences||Kihara Institute for Biological Research, Yokohama City UniversityCore Facility of Cell biology, Shanghai Center for Plant Stress Biology
分子生物学细胞生物学植物学
Kosami Ken-ichi,Kawasaki Tsutomu,Nagawa Shingo,Shimamoto Ko,Wong Hann Ling,Wang Qiong,Higuchi Masayuki,Inada Noriko,Okuda Jun,Matsuda Tomonori,Akamatsu Akira,Kawano Yoji,Tan Li.In vivo monitoring of plant small GTPase activation using a F?rster resonance energy transfer biosensor[EB/OL].(2025-03-28)[2025-05-16].https://www.biorxiv.org/content/10.1101/249938.点此复制
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