Abstract
We isolated a new pea mutant that was selected on the basis of pale color and elongated internodes in a screen under white light. The mutant was designated pcd1 for phytochrome chromophore deficient. Light-grown pcd1 plants have yellow-green foliage with a reduced chlorophyll (Chl) content and an abnormally high Chl a/Chl b ratio. Etiolated pcd1 seedlings are developmentally insensitive to far-red light, show a reduced response to red light, and have no spectrophotometrically detectable phytochrome. The phytochrome A apoprotein is present at the wild-type level in etiolated pcd1 seedlings but is not depleted by red light treatment. Crude phytochrome preparations from etiolated pcd1 tissue also lack spectral activity but can be assembled with phycocyanobilin, an analog of the endogenous phytochrome chromophore phytochromobilin, to yield a difference spectrum characteristic of an apophytochrome-phycocyanobilin adduct. These results indicate that the pcd1-conferred phenotype results from a deficiency in phytochrome chromophore synthesis. Furthermore, etioplast preparations from pcd1 seedlings can metabolize biliverdin (BV) IX[alpha] but not heme to phytochromobilin, indicating that pcd1 plants are severely impaired in their ability to convert heme to BV IX[alpha]. This provides clear evidence that the conversion of heme to BV IX[alpha] is an enzymatic process in higher plants and that it is required for synthesis of the phytochrome chromophore and hence for normal photomorphogenesis.
Full Text
The Full Text of this article is available as a PDF (1.8 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Chereskin B. M., Castelfranco P. A. Effects of Iron and Oxygen on Chlorophyll Biosynthesis : II. OBSERVATIONS ON THE BIOSYNTHETIC PATHWAY IN ISOLATED ETIOCHLOROPLASTS. Plant Physiol. 1982 Jan;69(1):112–116. doi: 10.1104/pp.69.1.112. [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]
- Cornejo J., Beale S. I. Algal heme oxygenase from Cyanidium caldarium. Partial purification and fractionation into three required protein components. J Biol Chem. 1988 Aug 25;263(24):11915–11921. [PubMed] [Google Scholar]
- Cornejo J., Beale S. I., Terry M. J., Lagarias J. C. Phytochrome assembly. The structure and biological activity of 2(R),3(E)-phytochromobilin derived from phycobiliproteins. J Biol Chem. 1992 Jul 25;267(21):14790–14798. [PubMed] [Google Scholar]
- Devlin P. F., Rood S. B., Somers D. E., Quail P. H., Whitelam G. C. Photophysiology of the Elongated Internode (ein) Mutant of Brassica rapa: ein Mutant Lacks a Detectable Phytochrome B-Like Polypeptide. Plant Physiol. 1992 Nov;100(3):1442–1447. doi: 10.1104/pp.100.3.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elich T. D., Lagarias J. C. Phytochrome Chromophore Biosynthesis : Both 5-Aminolevulinic Acid and Biliverdin Overcome Inhibition by Gabaculine in Etiolated Avena sativa L. Seedlings. Plant Physiol. 1987 Jun;84(2):304–310. doi: 10.1104/pp.84.2.304. [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]
- 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]
- Li L., Lagarias J. C. Phytochrome assembly. Defining chromophore structural requirements for covalent attachment and photoreversibility. J Biol Chem. 1992 Sep 25;267(27):19204–19210. [PubMed] [Google Scholar]
- López-Juez E., Nagatani A., Tomizawa K., Deak M., Kern R., Kendrick R. E., Furuya M. The cucumber long hypocotyl mutant lacks a light-stable PHYB-like phytochrome. Plant Cell. 1992 Mar;4(3):241–251. [PMC free article] [PubMed] [Google Scholar]
- Maines M. D. Heme oxygenase: function, multiplicity, regulatory mechanisms, and clinical applications. FASEB J. 1988 Jul;2(10):2557–2568. [PubMed] [Google Scholar]
- McDonagh A. F., Palma L. A. Preparation and properties of crystalline biliverdin IX alpha. Simple methods for preparing isomerically homogeneous biliverdin and [14C[biliverdin by using 2,3-dichloro-5,6-dicyanobenzoquinone. Biochem J. 1980 Aug 1;189(2):193–208. doi: 10.1042/bj1890193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Millar A. J., Straume M., Chory J., Chua N. H., Kay S. A. The regulation of circadian period by phototransduction pathways in Arabidopsis. Science. 1995 Feb 24;267(5201):1163–1166. doi: 10.1126/science.7855596. [DOI] [PubMed] [Google Scholar]
- Nagatani A., Reed J. W., Chory J. Isolation and Initial Characterization of Arabidopsis Mutants That Are Deficient in Phytochrome A. Plant Physiol. 1993 May;102(1):269–277. doi: 10.1104/pp.102.1.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neuhaus G., Bowler C., Kern R., Chua N. H. Calcium/calmodulin-dependent and -independent phytochrome signal transduction pathways. Cell. 1993 Jun 4;73(5):937–952. doi: 10.1016/0092-8674(93)90272-r. [DOI] [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]
- Parks B. M., Quail P. H. hy8, a new class of arabidopsis long hypocotyl mutants deficient in functional phytochrome A. Plant Cell. 1993 Jan;5(1):39–48. doi: 10.1105/tpc.5.1.39. [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]
- 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]
- Smith A. G., Santana M. A., Wallace-Cook A. D., Roper J. M., Labbe-Bois R. Isolation of a cDNA encoding chloroplast ferrochelatase from Arabidopsis thaliana by functional complementation of a yeast mutant. J Biol Chem. 1994 May 6;269(18):13405–13413. [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., 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., Maines M. D., Lagarias J. C. Inactivation of phytochrome- and phycobiliprotein-chromophore precursors by rat liver biliverdin reductase. J Biol Chem. 1993 Dec 15;268(35):26099–26106. [PubMed] [Google Scholar]
- Terry M. J., McDowell M. T., Lagarias J. C. (3Z)- and (3E)-phytochromobilin are intermediates in the biosynthesis of the phytochrome chromophore. J Biol Chem. 1995 May 12;270(19):11111–11118. doi: 10.1074/jbc.270.19.11111. [DOI] [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]
- Thomas J., Weinstein J. D. Measurement of heme efflux and heme content in isolated developing chloroplasts. Plant Physiol. 1990 Nov;94(3):1414–1423. doi: 10.1104/pp.94.3.1414. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinstein J. D., Beale S. I. Separate physiological roles and subcellular compartments for two tetrapyrrole biosynthetic pathways in Euglena gracilis. J Biol Chem. 1983 Jun 10;258(11):6799–6807. [PubMed] [Google Scholar]
- Weller J. L., Nagatani A., Kendrick R. E., Murfet I. C., Reid J. B. New lv Mutants of Pea Are Deficient in Phytochrome B. Plant Physiol. 1995 Jun;108(2):525–532. doi: 10.1104/pp.108.2.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whitelam G. C., Johnson E., Peng J., Carol P., Anderson M. L., Cowl J. S., Harberd N. P. Phytochrome A null mutants of Arabidopsis display a wild-type phenotype in white light. Plant Cell. 1993 Jul;5(7):757–768. doi: 10.1105/tpc.5.7.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Tuinen A., Kerckhoffs L. H., Nagatani A., Kendrick R. E., Koornneef M. Far-red light-insensitive, phytochrome A-deficient mutants of tomato. Mol Gen Genet. 1995 Jan 20;246(2):133–141. doi: 10.1007/BF00294675. [DOI] [PubMed] [Google Scholar]