Phytochrome Induction Research Articles

The protein kinase CK2 is involved in regulation of circadian rhythms in Arabidopsis.

A wide range of processes in plants, including expression of certain genes, is regulated by endogenous circadian rhythms. The circadian clock-associated 1 (CCA1) and the late elongated hypocotyl (LHY) proteins have been shown to be closely associated with clock function in Arabidopsis thaliana. The protein kinase CK2 can interact with and phosphorylate CCA1, but its role in the regulation of the circadian clock remains unknown. Here we show that plants overexpressing CKB3, a regulatory subunit of CK2, display increased CK2 activity and shorter periods of rhythmic expression of CCA1 and LHY. CK2 is also able to interact with and phosphorylate LHY in vitro. Additionally, overexpression of CKB3 shortened the periods of four known circadian clock-controlled genes with different phase angles, demonstrating that many clock outputs are affected. This overexpression also reduced phytochrome induction of an Lhcb gene. Finally, we found that the photoperiodic flowering response, which is influenced by circadian rhythms, was diminished in the transgenic lines, and that the plants flowered earlier on both long-day and short-day photoperiods. These data demonstrate that CK2 is involved in regulation of the circadian clock in Arabidopsis.

Constitutive Expression of the CIRCADIAN CLOCK ASSOCIATED 1 ( CCA1) Gene Disrupts Circadian Rhythms and Suppresses Its Own Expression

The CIRCADIAN CLOCK ASSOCIATED 1 ( CCA1) gene encodes a MYB-related transcription factor involved in the phytochrome induction of a light-harvesting chlorophyll a/ b-protein ( Lhcb) gene. Expression of the CCA1 gene is transiently induced by phytochrome and oscillates with a circadian rhythm. Constitutive expression of CCA1 protein in transgenic plants abolished the circadian rhythm of several genes with dramatically different phases. These plants also had longer hypocotyls and delayed flowering, developmental processes regulated by light and the circadian clock. Furthermore, the expression of both endogenous CCA1 and the related LHY gene was suppressed. Our results suggest that CCA1 is a part of a feedback loop that is closely associated with the circadian clock in Arabidopsis.

A Myb-related transcription factor is involved in the phytochrome regulation of an Arabidopsis Lhcb gene.

We have isolated the gene for a protein designated CCA1. This protein can bind to a region of the promoter of an Arabidopsis light-harvesting chlorophyll a/b protein gene, Lhcb1*3, which is necessary for its regulation by phytochrome. The CCA1 protein interacted with two imperfect repeats in the Lhcb1*3 promoter, AAA/cAATCT, a sequence that is conserved in Lhcb genes. A region near the N terminus of CCA1, which has some homology to the repeated sequence found in the DNA binding domain of Myb proteins, is required for binding to the Lhcb1*3 promoter. Lines of transgenic Arabidopsis plants expressing antisense RNA for CCA1 showed reduced phytochrome induction of the endogenous Lhcb1*3 gene, whereas expression of another phytochrome-regulated gene, rbcS-1A, which encodes the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, was not affected. Thus, the CCA1 protein acts as a specific activator of Lhcb1*3 transcription in response to brief red illumination. The expression of CCA1 RNA was itself transiently increased when etiolated seedlings were transferred to light. We conclude that the CCA1 protein is a key element in the functioning of the phytochrome signal transduction pathway leading to increased transcription of this Lhcb gene in Arabidopsis.

UV-B Inhibition of Phytochrome-Mediated Anthocyanin Formation in Sinapis alba L. Cotyledons

An action spectrum was measured for ultraviolet (UV) radiation-induced damage to (inhibition of) phytochrome-induced anthocyanin formation in cotyledons of 40-hour-old Sinapis alba L. seedlings. The action spectrum showed maximum effectiveness in the 260 to 280 nanometer waveband with little effect above 295 nanometers. The damaging effect of UV could be photorepaired by subsequent exposure to sunlight or to long wavelength (360 nanometers) UV radiation. Because this form of damage is subject to photorepair (photoreactivation), it is probably due to the formation of pyrimidine dimers, and the results suggest that it would not be ecologically relevant even if there was an increase in solar UV due to a decrease in stratospheric ozone levels of about 30%. If a dark period of more than 1 hour is interspersed between the phytochrome induction and the UV irradiation, the inhibition of the phytochrome induction gradually decreases with increasing dark period.

Anesthetic Effects on Secondary Dormancy and Phytochrome Responses in Setaria faberi Seeds

Seeds of giant foxtail (Setaria faberi Herm.) entered secondary dormancy after pretreatment in H(2)O at 35 degrees C. Pretreatment in 0.1 m ethanol, or several other substances with anesthetic properties, prevented secondary dormancy induction. Pretreatment in 0.5 m ethanol inhibited germination in darkness, but germination could be stimulated by a red irradiation. Germination was initially insensitive to light. Two separate responses are indicated. The first, affected by a variety of substances and low (0.1 m or less) concentrations of ethanol, is related to anesthetic effects and prevention of secondary dormancy. The second, induction of response to red irradiation, is caused by 0.5 m ethanol and some closely related substances. The anesthetic effect is accomplished within the first 8 hours of imbibition while the phytochrome induction effect required treatment for more than 24 hours. Both responses were lost if the 35 degrees C imbibition began in H(2)O. Involvement of cell membranes is suggested in the prevention of secondary dormancy by anesthetics.

Distorted phytochrome action spectra in green plants

Red light absorption by photosynthetic pigments screens stem tissue of Phaseolus vulgaris L. seedlings to such an extent that the most effective wavelength for the phytochrome induction control of stem growth is approximately 628 nm. Screening is greater in the first internode than in the hypocotyl. The consequences are that, compared with etiolated seedlings, much higher irradiances are required for phytochrome control in green plants, and the efficiency of 660 nm light is markedly reduced. Even very high exposures at 660 nm may not achieve appreciable photoconversion of P r to P fr.