Bioluminescent Monitoring of Circadian Rhythms in Isolated Mesophyll Cells of Arabidopsis at Single-Cell Level

Shunji Nakamura; Tokitaka Oyama

doi:10.1007/978-1-0716-2473-9_31

Bioluminescent Monitoring of Circadian Rhythms in Isolated Mesophyll Cells of Arabidopsis at Single-Cell Level

Methods Mol Biol. 2022:2525:395-405. doi: 10.1007/978-1-0716-2473-9_31.

Authors

Shunji Nakamura¹, Tokitaka Oyama²

Affiliations

¹ Department of Botany, Graduate School of Science, Kyoto University, Kitashirakawa-oiwake-cho, Kyoto, Japan.
² Department of Botany, Graduate School of Science, Kyoto University, Kitashirakawa-oiwake-cho, Kyoto, Japan. oyama.tokitaka.8w@kyoto-u.ac.jp.

PMID: 35836086
DOI: 10.1007/978-1-0716-2473-9_31

Abstract

A bioluminescent monitoring system is used to detect the circadian rhythms of individual plant cells. Transgenic Arabidopsis carrying the firefly luciferase (FLuc) gene driven by a circadian-regulated promoter is used as the material for protoplast isolation. The bioluminescence of these protoplasts in the culture medium is separately captured using a highly sensitive camera system. The time-series data of the bioluminescent imaging reveals the circadian rhythms of these isolated cells, enabling the native properties of the cellular circadian clocks to become elucidated.

Keywords: Arabidopsis thaliana; Bioluminescence (BL); Circadian rhythm; Firefly luciferase (FLuc); Mesophyll protoplast; Single cell.

MeSH terms

Arabidopsis* / genetics
Circadian Clocks*
Circadian Rhythm / genetics
Mesophyll Cells
Protoplasts