Circadian clock mutants in Arabidopsis identified by luciferase imaging

Science

Volumn 267, Issue 5201, 1995, Pages 1161-1163

  Millar, Andrew J ^a,b   Carré, Isabelle A ^a   Strayer, Carl A ^a   Chua, Nam Hai ^b   Kay, Steve A ^a  

^a UNIVERSITY OF VIRGINIA   (United States)

^b ROCKEFELLER UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

BINDING PROTEIN; CHLOROPHYLL; LUCIFERASE;

ARABIDOPSIS; ARTICLE; BIOLUMINESCENCE; CHROMOSOME 5; CIRCADIAN RHYTHM; GENE MUTATION; PHENOTYPE; PLANT LEAF; PLEIOTROPY; PRIORITY JOURNAL;

ARABIDOPSIS; ARABIDOPSIS SPP.;

EID: 0028902249     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.7855595     Document Type: Article

References (27)

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2. J. C. Hall, Annu. Rev. Genet. 24, 659 (1990); J. C. Dunlap, Annu. Rev. Physiol. 55, 683 (1983).
3. G. R. Teakle and S. A. Kay, unpublished results.
4. A. J. Millar, S. R. Short, N.-H. Chua, S. A. Kay, Plant Cell 4, 1075 (1992); A. J. Millar, S. R. Short, K. Hiratsuka, N.-H. Chua, S. A. Kay, Plant Mol. Biol. Rep. 10, 324 (1992).
5. A. J. Millar, S. R. Short, N.-H. Chua, S. A. Kay, Plant Cell 4, 1075 (1992); A. J. Millar, S. R. Short, K. Hiratsuka, N.-H. Chua, S. A. Kay, Plant Mol. Biol. Rep. 10, 324 (1992).
6. Seed homozygous for the cab2::Ω::Luc construct were soaked in 0.4% ethylmethane sulfonate for 12 hours at room temperature and sown in soil (M1 seed). Approximately 4500 M1 plants flowered. Self-fertilized seed (M2 seed) were harvested from pools of approximately 60 M1 plants.
7. The luminescence from 200 M2 seedlings from each of 39 M1 pools (total of ∼8000 M2 seedlings) was assayed by video imaging. Images were recorded when luminescence began to rise (19.5 hours after transfer to LL), close to the peak of luminescence (27.5 hours), and again near the trough (37 hours), when luminescence in control seedlings had returned to the level at 19.5 hours. Seedlings were returned to LL after each image.
8. We rescreened 250 candidate mutants by imaging every 3 hours over one LD cycle and 48 hours of LL.
9. A. J. Millar and S. A. Kay, unpublished results.
10. A. J. Millar, R. B. McGrath, N.-H. Chua, Annu. Rev. Genet. 28, 325 (1994).
11. Period estimation was performed by nonlinear least squares fitting of a modified sine wave, and variance-weighted means and standard deviations were calculated as described (10).
12. A. J. Millar, M. Straume, J. Chory, N.-H. Chua, S. A. Kay, Science 267, 1163 (1995).
13. Seedlings were grown for 7 days, in constant darkness or in LD. Hypocotyl length was measured on millimeter graph paper.
14. Ecotype Columbia-1 was provided by G. P. Rédei (University of Missouri-Columbia).
15. A. Konieczny and F. M. Ausubel, Plant J. 4, 403 (1993); C. J. Bell and J. R. Ecker, Genomics 19, 137 (1994); R. S. Reiter et al., Proc. Natl. Acad. Sci. U.S.A. 89, 1477 (1992). CAPS, cleaved amplified polymorphic sequences; RAPD, random amplified polymorphic DNA; SSLP, simple-sequence length polymorphism.
16. A. Konieczny and F. M. Ausubel, Plant J. 4, 403 (1993); C. J. Bell and J. R. Ecker, Genomics 19, 137 (1994); R. S. Reiter et al., Proc. Natl. Acad. Sci. U.S.A. 89, 1477 (1992). CAPS, cleaved amplified polymorphic sequences; RAPD, random amplified polymorphic DNA; SSLP, simple-sequence length polymorphism.
17. A. Konieczny and F. M. Ausubel, Plant J. 4, 403 (1993); C. J. Bell and J. R. Ecker, Genomics 19, 137 (1994); R. S. Reiter et al., Proc. Natl. Acad. Sci. U.S.A. 89, 1477 (1992). CAPS, cleaved amplified polymorphic sequences; RAPD, random amplified polymorphic DNA; SSLP, simple-sequence length polymorphism.
18. The toc1 gene showed linkage to CAPS marker DFR [50 recombinant chromosomes of 200 tested; recombination percentage (r) = 25.0 ± 3.1%, map distance by Kosambi's function (D) = 27.5 ± 4.1 centimorgans (cM)] and RAPD marker r488.2 (for one recombinant plant of 103 tested; r = 9.9 ± 2.1%, D = 10.0 ± 2.1 cM) [M. Koorneef and P. Stam, in Methods in Arabidopsis Research, C. Koncz, N.-H. Chua, J. Schell, Eds. (World Scientific, Singapore, 1992), pp. 83-99].
19. The transgene was mapped to chromosome 4 between the markers AG and PG11 in an independent cross between the transgenic line and Columbia (7). DET1 also maps to chromosome 4 [A. Pepper, T. Delaney, T. Washburn, D. Poole, J. Chory, Cell 78, 109 (1994)]; DET2 and COP1 map to chromosome 2 [J. Chory, P. Nagpal, C. Peto, Plant Cell 3, 445 (1991); X.-W. Deng and P. Quail, Plant J. 2, 83 (1992)].
20. The transgene was mapped to chromosome 4 between the markers AG and PG11 in an independent cross between the transgenic line and Columbia (7). DET1 also maps to chromosome 4 [A. Pepper, T. Delaney, T. Washburn, D. Poole, J. Chory, Cell 78, 109 (1994)]; DET2 and COP1 map to chromosome 2 [J. Chory, P. Nagpal, C. Peto, Plant Cell 3, 445 (1991); X.-W. Deng and P. Quail, Plant J. 2, 83 (1992)].
21. The transgene was mapped to chromosome 4 between the markers AG and PG11 in an independent cross between the transgenic line and Columbia (7). DET1 also maps to chromosome 4 [A. Pepper, T. Delaney, T. Washburn, D. Poole, J. Chory, Cell 78, 109 (1994)]; DET2 and COP1 map to chromosome 2 [J. Chory, P. Nagpal, C. Peto, Plant Cell 3, 445 (1991); X.-W. Deng and P. Quail, Plant J. 2, 83 (1992)].
22. A. J. Millar, Ph.D. thesis, Rockefeller University (1994).
23. G. Deitzer, in Light and the Flowering Process, D. Vince-Prue, B. Thomas, K. E. Cockshull, Eds. (Academic Press, New York, 1984), pp. 51-63.
24. C. R. McClung and S. A. Kay, in Arabidopsis thaliana, C. S. Somerville and E. Meyerowitz, Eds. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 615-637.
25. W. Engelmann, K. Simon, C. J. Phen, Z. NaturforSch. 47c, 925 (1992).
26. 3 family (see Fig. 2) were grown in LD for 17 days, then placed in a temperature-controlled incubator at 22°C. Interrupted lighting conditions simulated luminescence imaging protocols (2.5 hours light: 0.5 hour darkness). The OXALIS imaging system (19) recorded positions of true leaves every 10 min, from eight wild-type and eight mutant plants. Period estimates were calculated as described (10).
27. We thank K. W. Smith and D. Gerber for technical assistance, M. Straume and T. M. Breeden for programming, and D. Somers for critical reading of the manuscript. Supported by grants from the NSF Center for Biological Timing to S.A.K. and NIH grant GM44640 to N.-H.C. A.J.M. was supported by a William O. Baker Fellowship, through a grant from the Mellon Foundation. S.A.K. is supported by an award from the W. M. Keck Foundation.