Skip to main content
The Plant Cell logoLink to The Plant Cell
. 1991 Dec;3(12):1275–1288. doi: 10.1105/tpc.3.12.1275

Overexpression of Phytochrome B Induces a Short Hypocotyl Phenotype in Transgenic Arabidopsis.

D Wagner 1, JM Tepperman 1, PH Quail 1
PMCID: PMC160091  PMID: 12324591

Abstract

The photoreceptor phytochrome is encoded by a small multigene family in higher plants. phyA encodes the well-characterized etiolated-tissue phytochrome. The product of the phyB gene, which has properties resembling those of "green tissue" phytochrome, is as yet poorly characterized. We have developed a phytochrome B overexpression system for analysis of the structure and function of this protein. Using newly generated polyclonal and monoclonal antibodies that are selective for phytochrome B, we have demonstrated high levels of expression of full-length rice and Arabidopsis phytochrome B under the control of the cauliflower mosaic virus 35S promoter in transgenic Arabidopsis. The overexpressed phytochrome is spectrally active, undergoes red/far-red-light-dependent conformational changes, is synthesized in its inactive red light-absorbing form, and is stable in the light. Overexpression of phytochrome B is tightly correlated with a short hypocotyl phenotype in transgenic seedlings. This phenotype is strictly light dependent, thus providing direct evidence that phytochrome B is a biologically functional photoreceptor. Based on similarities to phenotypes obtained by overexpression of phytochrome A, it appears that phytochromes A and B can control similar responses in the plant.

Full Text

The Full Text of this article is available as a PDF (4.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adamse P., Jaspers P. A., Bakker J. A., Kendrick R. E., Koornneef M. Photophysiology and phytochrome content of long-hypocotyl mutant and wild-type cucumber seedlings. Plant Physiol. 1988 May;87(1):264–268. doi: 10.1104/pp.87.1.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boylan M. T., Quail P. H. Oat Phytochrome Is Biologically Active in Transgenic Tomatoes. Plant Cell. 1989 Aug;1(8):765–773. doi: 10.1105/tpc.1.8.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Cherry J. R., Hershey H. P., Vierstra R. D. Characterization of Tobacco Expressing Functional Oat Phytochrome : Domains Responsible for the Rapid Degradation of Pfr Are Conserved between Monocots and Dicots. Plant Physiol. 1991 Jul;96(3):775–785. doi: 10.1104/pp.96.3.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cleveland D. W. Peptide mapping in one dimension by limited proteolysis of sodium dodecyl sulfate-solubilized proteins. Methods Enzymol. 1983;96:222–229. doi: 10.1016/s0076-6879(83)96020-2. [DOI] [PubMed] [Google Scholar]
  6. Furuya M. Molecular properties and biogenesis of phytochrome I and II. Adv Biophys. 1989;25:133–167. doi: 10.1016/0065-227x(89)90006-3. [DOI] [PubMed] [Google Scholar]
  7. Hershey H. P., Barker R. F., Idler K. B., Lissemore J. L., Quail P. H. Analysis of cloned cDNA and genomic sequences for phytochrome: complete amino acid sequences for two gene products expressed in etiolated Avena. Nucleic Acids Res. 1985 Dec 9;13(23):8543–8559. doi: 10.1093/nar/13.23.8543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Karu A. E., Miller P. L., Chase C., Cornutt W. A point-addressable transfer system for automated sampling, feeding, and expansion of hybridoma cultures. J Immunol Methods. 1985 Jan 21;76(1):145–156. doi: 10.1016/0022-1759(85)90487-9. [DOI] [PubMed] [Google Scholar]
  9. Kay S. A., Nagatani A., Keith B., Deak M., Furuya M., Chua N. H. Rice Phytochrome Is Biologically Active in Transgenic Tobacco. Plant Cell. 1989 Aug;1(8):775–782. doi: 10.1105/tpc.1.8.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Keller J. M., Shanklin J., Vierstra R. D., Hershey H. P. Expression of a functional monocotyledonous phytochrome in transgenic tobacco. EMBO J. 1989 Apr;8(4):1005–1012. doi: 10.1002/j.1460-2075.1989.tb03467.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kende H., Lang A. Gibberellins and Light Inhibition of Stem Growth in Peas. Plant Physiol. 1964 May;39(3):435–440. doi: 10.1104/pp.39.3.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lagarias J. C., Lagarias D. M. Self-assembly of synthetic phytochrome holoprotein in vitro. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5778–5780. doi: 10.1073/pnas.86.15.5778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lagarias J. C., Mercurio F. M. Structure function studies on phytochrome. Identification of light-induced conformational changes in 124-kDa Avena phytochrome in vitro. J Biol Chem. 1985 Feb 25;260(4):2415–2423. [PubMed] [Google Scholar]
  14. Lee C., Levin A., Branton D. Copper staining: a five-minute protein stain for sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1987 Nov 1;166(2):308–312. doi: 10.1016/0003-2697(87)90579-3. [DOI] [PubMed] [Google Scholar]
  15. Nagai K., Thøgersen H. C. Synthesis and sequence-specific proteolysis of hybrid proteins produced in Escherichia coli. Methods Enzymol. 1987;153:461–481. doi: 10.1016/0076-6879(87)53072-5. [DOI] [PubMed] [Google Scholar]
  16. Quail P. H., Schäfer E., Marmé D. De novo synthesis of phytochrome in pumpkin hooks. Plant Physiol. 1973 Aug;52(2):124–127. doi: 10.1104/pp.52.2.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sharrock R. A., Quail P. H. Novel phytochrome sequences in Arabidopsis thaliana: structure, evolution, and differential expression of a plant regulatory photoreceptor family. Genes Dev. 1989 Nov;3(11):1745–1757. doi: 10.1101/gad.3.11.1745. [DOI] [PubMed] [Google Scholar]
  18. Somers D. E., Sharrock R. A., Tepperman J. M., Quail P. H. The hy3 Long Hypocotyl Mutant of Arabidopsis Is Deficient in Phytochrome B. Plant Cell. 1991 Dec;3(12):1263–1274. doi: 10.1105/tpc.3.12.1263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  20. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Valvekens D., Van Montagu M., Van Lijsebettens M. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536–5540. doi: 10.1073/pnas.85.15.5536. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES