Abstract

The plant circadian clock is an internal timekeeper that coordinates biological processes with daily changes in the external environment. The transcript levels of clock genes, which oscillate to control circadian outputs, were examined during early seedling development in barley (Hordeum vulgare), a model for temperate cereal crops. Oscillations of clock gene transcript levels do not occur in barley seedlings grown in darkness or constant light but were observed with day-night cycles. A dark-to-light transition influenced transcript levels of some clock genes but triggered only weak oscillations of gene expression, whereas a light-to-dark transition triggered robust oscillations. Single light pulses of 6, 12 or 18 hours induced robust oscillations. The light-to-dark transition was the primary determinant of the timing of subsequent peaks of clock gene expression. After the light-to-dark transition the timing of peak transcript levels of clock gene also varied depending on the length of the preceding light pulse. Thus, a single photoperiod can trigger initiation of photoperiod-dependent circadian rhythms in barley seedlings. Photoperiod-specific rhythms of clock gene expression were observed in two week old barley plants. Changing the timing of dusk altered clock gene expression patterns within a single day, showing that alteration of circadian oscillator behaviour is amongst the most rapid molecular responses to changing photoperiod in barley. A barley EARLY FLOWERING3 mutant, which exhibits rapid photoperiod–insensitive flowering behaviour, does not establish clock rhythms in response to a single photoperiod. The data presented show that dawn and dusk cues are important signals for setting the state of the circadian oscillator during early development of barley and that the circadian oscillator of barley exhibits photoperiod-dependent oscillation states.

Highlights

  • Plants, like most other organisms, exhibit rhythmic fluctuations in gene expression and metabolism synchronised with day-night cycles

  • Clock genes do not oscillate in seedlings germinated in constant conditions Clock gene transcript levels were first assayed in 5 day old barley seedlings grown in 12 hour day-night cycles

  • Rhythmic oscillations of transcript levels were observed for several clock genes including HvCCA1, HvTOC1, HvGI, HvPRR73, HvPRR59 and HvPRR95 (Fig 1A, S1 Fig), similar to previous reports (e.g. [24])

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Summary

Introduction

Like most other organisms, exhibit rhythmic fluctuations in gene expression and metabolism synchronised with day-night cycles. Genetic studies of model systems such as Arabidopsis (Arabidopsis thaliana) have identified components of the plant circadian clock These include the MYB transcription factor genes CIRCADIAN CLOCK ASSOCIATED1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY) and a series of PSEUDO RESPONSE REGULATOR (PRR) genes including TIMING OF CAB EXPRESSION 1 (TOC1) [4,5,6]. Transcript levels for these genes oscillate with regular daily peaks and troughs. Circadian clock genes form a molecular mechanism that regulates daily rhythms of clock outputs

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