Large-scale screening of Arabidopsis circadian clock mutants by a high-throughput real-time bioluminescence monitoring system.

Using a high-throughput real-time bioluminescence monitoring system, we screened large numbers of Arabidopsis thaliana mutants for extensively altered circadian rhythms. We constructed reporter genes by fusing a promoter of an Arabidopsis flowering-time gene - either GIGANTEA (GI) or FLOWERING LOCUS T (FT) - to a modified firefly luciferase gene (LUC(+)), and we transferred the fusion gene (P(GI)::LUC(+) or P(FT)::LUC(+)) into the Arabidopsis genome. After mutagenesis with ethyl methanesulfonate, 50 000 M(2) seedlings carrying the P(GI)::LUC(+) and 50 000 carrying P(FT)::LUC(+) were screened their bioluminescence rhythms. We isolated six arrhythmic (AR) mutants and 29 other mutants that showed more than 3 h difference in the period length or phase of rhythms compared with the wild-type strains. The shortest period length was 16 h, the longest 27 h. Five of the six AR mutants carrying P(GI)::LUC(+) showed arrhythmia in bioluminescence rhythms in both constant light and constant dark. These five AR mutants also showed arrhythmia in leaf movement rhythms in constant light. Genetic analysis revealed that each of the five AR mutants carried a recessive mutation in a nuclear gene and the mutations belonged to three complementation groups, and at least one of which was mapped on a novel locus. Our results suggest that the three loci identified here may contain central clock or clock-related genes, at least one of which may be a novel.

[1]  Ralf Stanewsky,et al.  Genetic analysis of the circadian system in Drosophila melanogaster and mammals. , 2003, Journal of neurobiology.

[2]  Joanna Putterill,et al.  The late elongated hypocotyl Mutation of Arabidopsis Disrupts Circadian Rhythms and the Photoperiodic Control of Flowering , 1998, Cell.

[3]  J. Dunlap.  Molecular Bases for Circadian Clocks , 1999, Cell.

[4]  Takeshi Mizuno,et al.  The APRR1/TOC1 quintet implicated in circadian rhythms of Arabidopsis thaliana: I. Characterization with APRR1-overexpressing plants. , 2002, Plant & cell physiology.

[5]  Serge Daan,et al.  A functional analysis of circadian pacemakers in nocturnal rodents , 1976, Journal of comparative physiology.

[6]  D. E. Somers,et al.  ZEITLUPE Encodes a Novel Clock-Associated PAS Protein from Arabidopsis , 2000, Cell.

[7]  C. R. McClung,et al.  The Arabidopsis Circadian System , 2002, The arabidopsis book.

[8]  J. Dunlap,et al.  Genetic and molecular analysis of circadian rhythms in Neurospora. , 2001, Annual review of physiology.

[9]  M. W. Young,et al.  Product of per locus of Drosophila shares homology with proteoglycans , 1986, Nature.

[10]  G. Coupland,et al.  GIGANTEA: a circadian clock‐controlled gene that regulates photoperiodic flowering in Arabidopsis and encodes a protein with several possible membrane‐spanning domains , 1999, The EMBO journal.

[11]  D. R. Wagner,et al.  EARLY FLOWERING3 Encodes a Novel Protein That Regulates Circadian Clock Function and Flowering in Arabidopsis , 2001, The Plant Cell Online.

[12]  M. Ishiura,et al.  Circadian Rhythms in the Thermophilic Cyanobacterium Thermosynechococcus elongatus: Compensation of Period Length over a Wide Temperature Range , 2004, Journal of bacteriology.

[13]  Tsuyoshi Mizoguchi,et al.  LHY and CCA1 are partially redundant genes required to maintain circadian rhythms in Arabidopsis. , 2002, Developmental cell.

[14]  E. Tobin,et al.  Loss of the circadian clock-associated protein 1 in Arabidopsis results in altered clock-regulated gene expression. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  S. Kay,et al.  Orchestrated transcription of key pathways in Arabidopsis by the circadian clock. , 2000, Science.

[16]  C. Strayer,et al.  Circadian clock mutants in Arabidopsis identified by luciferase imaging , 1995, Science.

[17]  Yoshiyuki Sakaki,et al.  Circadian oscillation of a mammalian homologue of the Drosophila period gene , 1997, Nature.

[18]  M. Bevan,et al.  GUS fusions: beta‐glucuronidase as a sensitive and versatile gene fusion marker in higher plants. , 1987, The EMBO journal.

[19]  F Halberg,et al.  Methods for cosinor-rhythmometry. , 1979, Chronobiologia.

[20]  Y. Kobayashi,et al.  A pair of related genes with antagonistic roles in mediating flowering signals. , 1999, Science.

[21]  C Robertson McClung,et al.  CIRCADIAN RHYTHMS IN PLANTS. , 2003, Annual review of plant physiology and plant molecular biology.

[22]  J. Chory,et al.  The Arabidopsis SRR1 gene mediates phyB signaling and is required for normal circadian clock function. , 2003, Genes & development.

[23]  Stacey L. Harmer,et al.  Critical Role for CCA1 and LHY in Maintaining Circadian Rhythmicity in Arabidopsis , 2002, Current Biology.

[24]  J. Glazebrook,et al.  Arabidopsis : a laboratory manual , 2002 .

[25]  Michael F. Covington,et al.  ELF3 Modulates Resetting of the Circadian Clock in Arabidopsis , 2001, The Plant Cell Online.

[26]  D. E. Somers,et al.  Cloning of the Arabidopsis clock gene TOC1, an autoregulatory response regulator homolog. , 2000, Science.

[27]  J. W. Hastings,et al.  Acquisition of Circadian Bioluminescence Data in Gonyaulax and an Effect of the Measurement Procedure on the Period of the Rhythm , 1986, Journal of biological rhythms.

[28]  A. Millar.  Biological clocks in Arabidopsis thaliana. , 1999, The New phytologist.

[29]  Gregor Eichele,et al.  RIGUI, a Putative Mammalian Ortholog of the Drosophila period Gene , 1997, Cell.

[30]  Anthony Hall,et al.  The TIME FOR COFFEE Gene Maintains the Amplitude and Timing of Arabidopsis Circadian Clocks Online version contains Web-only data. Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.013730. , 2003, The Plant Cell Online.

[31]  T. Mizuno,et al.  Genes encoding pseudo-response regulators: insight into His-to-Asp phosphorelay and circadian rhythm in Arabidopsis thaliana. , 2000, Plant & cell physiology.

[32]  A. Millar,et al.  Circadian dysfunction causes aberrant hypocotyl elongation patterns in Arabidopsis. , 1999, The Plant journal : for cell and molecular biology.

[33]  C. R. McClung,et al.  The out of phase 1 Mutant Defines a Role for PHYB in Circadian Phase Control in Arabidopsis1 , 2002, Plant Physiology.

[34]  C. Koncz,et al.  The promoter of TL-DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type of Agrobacterium binary vector , 1986, Molecular and General Genetics MGG.

[35]  D. E. Somers,et al.  Conditional Circadian Dysfunction of the Arabidopsis early-flowering 3 Mutant , 1996, Science.

[36]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .

[37]  D. E. Somers,et al.  Control of circadian rhythms and photoperiodic flowering by the Arabidopsis GIGANTEA gene. , 1999, Science.

[38]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[39]  Andrew J. Millar,et al.  Biological rhythms and photoperiodism in plants , 1998 .

[40]  S. Kay,et al.  Living by the calendar: how plants know when to flower , 2003, Nature Reviews Molecular Cell Biology.

[41]  J. Chory,et al.  Activation tagging of the floral inducer FT. , 1999, Science.

[42]  Zhi-Yong Wang,et al.  Constitutive Expression of the CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) Gene Disrupts Circadian Rhythms and Suppresses Its Own Expression , 1998, Cell.

[43]  E. Huq,et al.  GIGANTEA is a nuclear protein involved in phytochrome signaling in Arabidopsis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[44]  D. E. Somers,et al.  The short-period mutant, toc1-1, alters circadian clock regulation of multiple outputs throughout development in Arabidopsis thaliana. , 1998, Development.

[45]  Hitoshi Onouchi,et al.  CONSTANS mediates between the circadian clock and the control of flowering in Arabidopsis , 2001, Nature.

[46]  G. Coupland,et al.  Shedding light on the circadian clock and the photoperiodic control of flowering. , 2003, Current opinion in plant biology.

[47]  Steve A. Kay,et al.  Reciprocal Regulation Between TOC1 and LHY/CCA1 Within the Arabidopsis Circadian Clock , 2001, Science.

[48]  C. Johnson,et al.  Expression of a gene cluster kaiABC as a circadian feedback process in cyanobacteria. , 1998, Science.

[49]  C. R. McClung,et al.  The Neurospora clock gene frequency shares a sequence element with the Drosophila clock gene period , 1989, Nature.

[50]  Andrew J. Millar,et al.  The ELF4 gene controls circadian rhythms and flowering time in Arabidopsis thaliana , 2002, Nature.