期刊
PLANT AND CELL PHYSIOLOGY
卷 54, 期 12, 页码 2085-2093出版社
OXFORD UNIV PRESS
DOI: 10.1093/pcp/pct131
关键词
Bioluminescence; Gene expression; Lemna gibba; Luciferase; Particle bombardment; Single cells
资金
- Japan Society for the Promotion of Science (JSPS) KAKENHI [23657033, 25650098, 24-1530]
- Iwatani Naoji Foundation
- Japan Science and Technology Agency (JST)
- Grants-in-Aid for Scientific Research [25650098, 23657033] Funding Source: KAKEN
Gene expression is a fundamental cellular process and expression dynamics are of great interest in life science. We succeeded in monitoring cellular gene expression in a duckweed plant, Lemna gibba, using bioluminescent reporters. Using particle bombardment, epidermal and mesophyll cells were transfected with the luciferase gene (luc+) under the control of a constitutive [Cauliflower mosaic virus 35S (CaMV35S)] and a rhythmic [Arabidopsis thaliana CIRCADIAN CLOCK ASSOCIATED 1 (AtCCA1)] promoter. Bioluminescence images were captured using an EM-CCD (electron multiply charged couple device) camera. Luminescent spots of the transfected cells in the plant body were quantitatively measured at the single-cell level. Luminescence intensities varied over a 1,000-fold range among CaMV35S::luc+-transfected cells in the same plant body and showed a log-normal-like frequency distribution. We monitored cellular gene expression under light-dark conditions by capturing bioluminescence images every hour. Luminescence traces of >= 50 individual cells in a frond were successfully obtained in each monitoring procedure. Rhythmic and constitutive luminescence behaviors were observed in cells transfected with AtCCA1::luc+ and CaMV35S::luc+, respectively. Diurnal rhythms were observed in every AtCCA1::luc+-introduced cell with traceable luminescence, and slight differences were detected in their rhythmic waveforms. Thus the single-cell bioluminescence monitoring system was useful for the characterization of cellular gene expression in a plant body.
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