Abstract
The photoregulatory activity of the phytochrome photoreceptor requires the synthesis and covalent attachment of the linear tetrapyrrole prosthetic group phytochromobilin. Because the mammalian enzyme biliverdin IX alpha reductase (BVR) is able to functionally inactivate phytochromobilin in vitro, this investigation was undertaken to determine whether BVR expression in transgenic plants would prevent the synthesis of functionally active phytochrome in vivo. Here, we show that plastid-targeted, constitutive expression of BVR in Arabidopsis yields plants that display aberrant photomorphogenesis throughout their life cycle. Photobiological and biochemical analyses of three transgenic BVR lines exhibiting a 25-fold range of BVR expression established that the BVR-dependent phenotypes are light dependent, pleiotropic, and consonant with the loss of multiple phytochrome activities. Chlorophyll accumulation in BVR-expressing transgenic plants was particularly sensitive to increased light fluence rates, which is consistent with an important role for phytochrome in light tolerance. Under blue light, transgenic BVR plants displayed elongated hypocotyls but retained phototropic behavior and the ability to fully deetiolate. Directed BVR expression may prove to be useful for probing the cellular and developmental basis of phytochrome-mediated responses and for selective control of individual aspects of light-mediated plant growth and development.
Full Text
The Full Text of this article is available as a PDF (3.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ahmad M., Cashmore A. R. HY4 gene of A. thaliana encodes a protein with characteristics of a blue-light photoreceptor. Nature. 1993 Nov 11;366(6451):162–166. doi: 10.1038/366162a0. [DOI] [PubMed] [Google Scholar]
- Beale S. I. Biosynthesis of the Tetrapyrrole Pigment Precursor, delta-Aminolevulinic Acid, from Glutamate. Plant Physiol. 1990 Aug;93(4):1273–1279. doi: 10.1104/pp.93.4.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Becker D. Binary vectors which allow the exchange of plant selectable markers and reporter genes. Nucleic Acids Res. 1990 Jan 11;18(1):203–203. doi: 10.1093/nar/18.1.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berry-Lowe S. L., Mc Knight T. D., Shah D. M., Meagher R. B. The nucleotide sequence, expression, and evolution of one member of a multigene family encoding the small subunit of ribulose-1,5-bisphosphate carboxylase in soybean. J Mol Appl Genet. 1982;1(6):483–498. [PubMed] [Google Scholar]
- 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]
- Carrington J. C., Freed D. D., Oh C. S. Expression of potyviral polyproteins in transgenic plants reveals three proteolytic activities required for complete processing. EMBO J. 1990 May;9(5):1347–1353. doi: 10.1002/j.1460-2075.1990.tb08249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chory J., Peto C. A., Ashbaugh M., Saganich R., Pratt L., Ausubel F. Different Roles for Phytochrome in Etiolated and Green Plants Deduced from Characterization of Arabidopsis thaliana Mutants. Plant Cell. 1989 Sep;1(9):867–880. doi: 10.1105/tpc.1.9.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elich T. D., McDonagh A. F., Palma L. A., Lagarias J. C. Phytochrome chromophore biosynthesis. Treatment of tetrapyrrole-deficient Avena explants with natural and non-natural bilatrienes leads to formation of spectrally active holoproteins. J Biol Chem. 1989 Jan 5;264(1):183–189. [PubMed] [Google Scholar]
- Gaffney T., Friedrich L., Vernooij B., Negrotto D., Nye G., Uknes S., Ward E., Kessmann H., Ryals J. Requirement of salicylic Acid for the induction of systemic acquired resistance. Science. 1993 Aug 6;261(5122):754–756. doi: 10.1126/science.261.5122.754. [DOI] [PubMed] [Google Scholar]
- Inskeep W. P., Bloom P. R. Extinction coefficients of chlorophyll a and B in n,n-dimethylformamide and 80% acetone. Plant Physiol. 1985 Feb;77(2):483–485. doi: 10.1104/pp.77.2.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jabben M., Shanklin J., Vierstra R. D. Red light-induced accumulation of ubiquitin-phytochrome conjugates in both monocots and dicots. Plant Physiol. 1989 Jun;90(2):380–384. doi: 10.1104/pp.90.2.380. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karlin-Neumann G. A., Sun L., Tobin E. M. Expression of Light-Harvesting Chlorophyll a/b-Protein Genes Is Phytochrome-Regulated in Etiolated Arabidopsis thaliana Seedlings. Plant Physiol. 1988 Dec;88(4):1323–1331. doi: 10.1104/pp.88.4.1323. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kraepiel Y., Jullien M., Cordonnier-Pratt M. M., Pratt L. Identification of two loci involved in phytochrome expression in Nicotiana plumbaginifolia and lethality of the corresponding double mutant. Mol Gen Genet. 1994 Mar;242(5):559–565. doi: 10.1007/BF00285279. [DOI] [PubMed] [Google Scholar]
- Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Liscum E., Briggs W. R. Mutations in the NPH1 locus of Arabidopsis disrupt the perception of phototropic stimuli. Plant Cell. 1995 Apr;7(4):473–485. doi: 10.1105/tpc.7.4.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liscum E., Hangarter R. P. Arabidopsis Mutants Lacking Blue Light-Dependent Inhibition of Hypocotyl Elongation. Plant Cell. 1991 Jul;3(7):685–694. doi: 10.1105/tpc.3.7.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parks B. M., Quail P. H. Phytochrome-Deficient hy1 and hy2 Long Hypocotyl Mutants of Arabidopsis Are Defective in Phytochrome Chromophore Biosynthesis. Plant Cell. 1991 Nov;3(11):1177–1186. doi: 10.1105/tpc.3.11.1177. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quail P. H., Boylan M. T., Parks B. M., Short T. W., Xu Y., Wagner D. Phytochromes: photosensory perception and signal transduction. Science. 1995 May 5;268(5211):675–680. doi: 10.1126/science.7732376. [DOI] [PubMed] [Google Scholar]
- Quail P. H. Phytochrome: a light-activated molecular switch that regulates plant gene expression. Annu Rev Genet. 1991;25:389–409. doi: 10.1146/annurev.ge.25.120191.002133. [DOI] [PubMed] [Google Scholar]
- Quail P. H., Schäfer E., Marmé D. Turnover of phytochrome in pumpkin cotyledons. Plant Physiol. 1973 Aug;52(2):128–131. doi: 10.1104/pp.52.2.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reed J. W., Nagatani A., Elich T. D., Fagan M., Chory J. Phytochrome A and Phytochrome B Have Overlapping but Distinct Functions in Arabidopsis Development. Plant Physiol. 1994 Apr;104(4):1139–1149. doi: 10.1104/pp.104.4.1139. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reinbothe S., Reinbothe C. Regulation of Chlorophyll Biosynthesis in Angiosperms. Plant Physiol. 1996 May;111(1):1–7. doi: 10.1104/pp.111.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shinomura T., Nagatani A., Hanzawa H., Kubota M., Watanabe M., Furuya M. Action spectra for phytochrome A- and B-specific photoinduction of seed germination in Arabidopsis thaliana. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):8129–8133. doi: 10.1073/pnas.93.15.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
- 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]
- Terry M. J., Kendrick R. E. The aurea and yellow-green-2 mutants of tomato are deficient in phytochrome chromophore synthesis. J Biol Chem. 1996 Aug 30;271(35):21681–21686. doi: 10.1074/jbc.271.35.21681. [DOI] [PubMed] [Google Scholar]
- Terry M. J., Lagarias J. C. Holophytochrome assembly. Coupled assay for phytochromobilin synthase in organello. J Biol Chem. 1991 Nov 25;266(33):22215–22221. [PubMed] [Google Scholar]
- Terry M. J., Wahleithner J. A., Lagarias J. C. Biosynthesis of the plant photoreceptor phytochrome. Arch Biochem Biophys. 1993 Oct;306(1):1–15. doi: 10.1006/abbi.1993.1473. [DOI] [PubMed] [Google Scholar]
- 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]
- Weller J. L., Terry M. J., Rameau C., Reid J. B., Kendrick R. E. The Phytochrome-Deficient pcd1 Mutant of Pea Is Unable to Convert Heme to Biliverdin IX[alpha]. Plant Cell. 1996 Jan;8(1):55–67. doi: 10.1105/tpc.8.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wessel D., Flügge U. I. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem. 1984 Apr;138(1):141–143. doi: 10.1016/0003-2697(84)90782-6. [DOI] [PubMed] [Google Scholar]