Transcription of Arabidopsis and wheat Cab genes in single tobacco transgenic seedlings exhibits independent rhythms in a developmentally regulated fashion

Abstract

Transcription of Cab genes has been previously shown to be regulated by a circadian oscillator coupled to the red light-absorbing plant photoreceptor phytochrome in various plant species. In addition, it has recently been suggested that rhythmic expression of the Cab genes could also be affected by a phytochrome-independent circadian oscillator in a developmentally regulated fashion. This study has shown that a red light-insensitive oscillator and a phytochrome-coupled circadian clock indeed coregulate the oscillating expression of individual Cab genes at the level of transcription at an early developmental stage. The study involved analysing the expression patterns of transgenes, containing short fragments of the Arabidopsis thaliana Cab2 or the wheat Cab-1 promoter fused to the firefly luciferase reporter gene, by a video-imaging system in single, etiolated tobacco seedlings. Germination and red/far-red light treatments applied between 12 and 36 h after sowing lead to the appearance of two independent circadian rhythms. These rhythms co-exist, both exhibiting period lengths close to 25 h but phased differently. However, repeated red-light treatments given 60 h or later after sowing synchronize these free-running rhythms and induce a single new circadian oscillation. These data indicate that both oscillators regulate the expression of the Cab genes studied at the level of transcription and that the cis-acting element(s) of the wheat Cab-1 and A. thaliana Cab2 genes mediating these responses are located on short, 250 bp promoter regions. Furthermore, these red-light induced rhythms are also inducible by far-red light treatments alone. Therefore, in tobacco, the phytochrome-coupled oscillator is regulated, at least partially, by the very low fluence response of phytochrome A.