Model-based analysis of the circadian rhythm generation of bioluminescence reporter activity in duckweed.

Bioluminescence monitoring techniques are widely used to study the gene expression dynamics in living plants. Monitoring the bioluminescence from a luciferase gene under the control of a circadian promoter is indispensable for examining plant circadian systems. The bioluminescence monitoring technique was successfully applied to physiological studies of circadian rhythms in duckweed plants. It has been reported that a luciferase gene under a constitutive promoter also exhibits a bioluminescent circadian rhythm in duckweed. However, the mechanisms underlying rhythm generation remain unknown. In this study, we performed a model-based analysis to evaluate the machinery that generates the bioluminescence rhythm. We hypothesized the rhythmic factor of three aspects regarding the bioluminescence intensities of luciferase in cells: luminescence efficiency, production rate, and degradation rate. Theoretically, if the latter two are involved in rhythm generation, the difference in luciferase stability affects the amplitude and phase relations of the bioluminescence rhythm. Luciferase stability is irrelevant to these rhythm properties if only the luminescence efficiency is involved. First, we simulated the bioluminescence rhythms of two luciferases with different stabilities associated with each of three rhythmic factors. Luciferase stability was set based on the reported values for Emerald-luciferase and Emerald-luciferase-PEST. We then experimentally examined the bioluminescence rhythms of reporters of these luciferases driven by the CAULIFLOWER MOSAIC VIRUS 35S promoter in the duckweed Lemna japonica. Their circadian properties matched those obtained from the simulation of the luminescence efficiency. This supports the view that cells in duckweed show circadian changes in physiological conditions associated with the luciferase enzyme reaction.