Abstract
We have isolated and sequenced a cDNA clone encoding the apoprotein of a potato phytochrome. Based on the deduced amino acid sequence, which shows 78% amino acid identity to the Arabidopsis phyA and 50% identity to the Arabidopsis phyB open reading frame, we have classified this cDNA clone as potato phyA phytochrome. The amino acid immediately preceding cysteine 323, which is the homologue of oat cystein 321, to which the chromophore has been shown to be attached, is a tyrosine residue. This contrasts with six other type A phytochrome sequences from both monocots and dicots that encode serine in this position. As already observed in three other cDNAs isolated from dicot species, the potato phyA clone encodes a short open reading frame (13 amino acids) preceding the phyA open reading frame (1123 amino acids), supporting the idea that this type of leader sequence might be involved in the regulated expression of the phytochrome apoprotein. Southern blot analysis revealed a single phyA gene as well as other related phytochrome sequences in the potato genome. phyA mRNA levels varied in different organs and were modulated by white light; in seedlings and sprouts, highest levels of mRNA were detected in the etiolated stage. Upon illumination with white light, mRNA levels decreased to the amount found in leaves of re-etiolated plants. Lowest expression was observed in leaves of plants grown in the light, in tubers irrespective of light treatment, and in roots of plants grown in the dark. In roots of plants grown in the light, elevated levels of phyA mRNA were detected. Using a monoclonal antibody generated against pea phytochrome as an immunochemical probe, the protein was only detectable in protein extracts from etiolated seedlings and sprouts.
Similar content being viewed by others
References
Balton GW, Quail PH: Cell-free synthesis of phytochrome apoprotein. Planta 155: 212–217 (1982).
Batutis EJ, Ewing EE: Far-red reversal of red light effect during long-night induction of potato (Solanum tuberosum L.) tuberization. Plant Physiol 69: 672–674 (1982).
Biggin MD, Gibson TJ, Hong GF: Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80: 3963–3965 (1983).
Christensen AH, Quail PH: Structure and expression of a maize phytochrome-encoding gene. Gene 85: 381–395 (1989).
Cordonnier MM, Greppin H, Pratt LH: Identification of a highly conserved domain on phytochrome from angiosperms to algae. Plant Physiol 80: 982–987 (1986).
Cordonnier M-M, Greppin H, Pratt LH: Phytochrome from green Avena shoots characterized with a monoclonal antibody to phytochrome from etiolated Pisum shoots. Biochemistry 25: 7657–7666 (1986).
Dehesh K, Tepperman J, Christensen AH, Quail PH: PhyB is evolutionarily conserved and constitutively expressed in rice seedling shoots. Mol Gen Genet 225: 305–313 (1991).
Devi KRG, Chan Y-L, Wool IG: The primary structure of rat ribosomal protein S4. Biochim Biophys Acta 1008: 258–262 (1989).
Eckes P, Schell J, Willmitzer L: Isolation and characterization of a light-inducible, organ-specific gene from potato and analysis of its expression after tagging and transfer into tobacco and potato shoots. Mol Gen Genet 205: 14–22 (1986).
Gatz C, Kaiser A, Wendenburg R: Regulation of a modified CaMV 35S promoter by the Tn10-encoded Tet repressor in transgenic tobacco. Mol Gen Genet 227: 229–237 (1991).
Grimm R, Rüdiger W: A simple and rapid method for isolation of 124 kDa oat phytochrome. Z Naturforsch 084 41c: 988–992 (1988).
Hershey HP, Barker RF, Idler KB, Lissemore JL, Quail PH: Analysis of cloned cDNA and genomic sequences for phytochrome: Amino acid sequences for two gene products expressed in etiolated Avena. Nucl Acids Res 13: 8543–8559 (1985).
Kay SA, Keith B, Shinozaki, Chye M-L, Chua N-H: The rice phytochrome gene: Structure, autoregulated expression, and binding of GT-1 to a conserved site in the 5′ upstream region. Plant Cell 1: 351–360 (1989).
Kay SA, Keith B, Shinozaki, Chua N-H: The sequence of the rice phytochrome gene. Nucl Acids Res 17: 2865–2866 (1989).
Keil M, Sanchez-Serrano JJ, Willmitzer L: Both wound inducible and tuber specific expression are mediated by the promoter of a single member of the potato proteinase inhibitor II gene family. EMBO J 8: 1323–1330 (1989).
Köster-Töpfer M, Frommer W, Rocha-Sosa M, Rosahl S, Schell J, Willmitzer L: A class II patatin promoter is under developmental control in both transgenic potato and tobacco plants. Mol Gen Genet 219: 390–396 (1989).
Kronenberg GHM, Kendrick RE: The physiology of action. In: Kronenberg GHM, Kendrick RE (eds) Photomorphogenesis in Plants, pp 99–114. Martinus Nijhoff Publishers, Dordrecht (1986).
Kyhse-Andersen J: Electroblotting of multiple gels: A simple apparatus without buffer-tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Meth 10: 203–209 (1984).
Lagarias JC, Rapoport H: Chromopeptides from phytochrome. The structure and linkage of the Pr form of the phytochrome chromophore. J Am Chem Soc 102: 4821–4828 (1980).
Lagarias JC: Progress in the molecular analysis of phytochrome. Photochem Photobiol 42: 811–820 (1985).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Lissemore JL, Colbert JT, quail PH: Cloning of cDNA for phytochrome from etiolated Cucurbita and coordinate photoregulation of the abundance of two distinct phytochrome transcripts. Plant Mol Biol 8: 485–496 (1987).
Logemann J, Schell J, Willmitzer L: Improved method for the isolation of RNA from plant tissues. Anal Biochem 163: 16–20 (1987).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1982).
Mohr H, Drumm-Herrel H, Oelmüller R: Coaction of phytochrome and blue/UV light photoreceptors. In: Senger H (ed) Blue Light Effects in Biological Systems, pp 6–19. Springer-Verlag, Heidelberg/Berlin (1984).
Morris SC, Graham D, Lee TH: Phytochrome control of chlorophyll synthesis in potato tubers. Plant Sci Lett 17: 13–19 (1979).
Mueller PP, Hinnebush AG: Multiple upstream AUG codons mediate translational control of GCN4. Cell 45: 201–207 (1986).
Murray NE: Phage lambda and molecular cloning. In: Hendrix RW, Roberts JW, Stahl FW, Weisberger RA (eds) Lambda II, pp. 395–432. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1983).
Rocha-Sosa M, Sonnewald U, Frommer W, Stratmann M, Schell J, Willmitzer L: Both developmental and metabolic signals activate the promoter of class I patatin gene. EMBO J 8: 23–29 (1989).
Rogers SO, Bendich AJ: Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol Biol 5: 69–76 (1985).
Rüdiger W, Scheer H: Chromophores in photomorphogenesis. In: Shropshire WJr, Mohr H (eds) Photomorphogenesis. Encyclopedia of Plant Physiology, New Series 16A, pp. 119–151. Springer-Verlag, Heidelberg/Berlin. (1983).
Rüdiger W: Biochemistry of the phytochrome chromophore. In: Furuya M (ed) Phytochrome and Photoregulation in Plants, pp. 127–137. Academic Press, New York (1987).
Salisbury FB, Ross CW: Plant physiology. Wadsworth Publishing Company, Belmont (1985).
Sato N: Nucleotide sequence and expression of the phytochrome gene in Pisum sativum: Differential regulation by light of multiple transcripts. Plant Mol Biol 11: 697–710 (1989).
Sharrock RA, Lissemore JL, Quail PH: Nucleotide and amino acid sequence of a Cucurbita phytochrome cDNA clone: Identification of conserved features by comparison with Avena phytochrome. Gene 47: 287–295 (1986).
Sharrock RA, Parks BM, Koorneef M, Quail PH: Molecular analysis of the phytochrome deficiency in an aurea mutant of tomato. Mol Gen Genet 213: 9–14 (1988).
Sharrock RA, Quail PH: Novel phytochrome sequences in Arabidopsis thaliana: structure, evolution and differential expression of a plant regulatory photoreceptor family. Gene Devel 3: 1745–1757 (1989).
Shropshire WJr, Mohr H (eds) Photomorphogenesis. Encyclopedia of Plant Physiology, New Series 16A, pp. 119–151. Springer-Verlag, Heidelberg/Berlin (1983).
Stockhaus J, Schell J, Willmitzer L: Correlation of the expression of the nuclear photosynthetic gene ST-LS1 with the presence of chloroplasts. EMBO J 8: 2445–2451 (1989).
Sullivan DS, Christensen AH, Quail PH: Isolation and characterization of a maize chlorophyll a/b binding protein gene that produces high levels of mRNA in the dark. Mol Gen Genet 215: 431–440 (1989).
Tokuhisa JG, Quail PH: Phytochrome in green tissue: spectral and immunochemical evidence for two distinct molecular species of phytochrome in light-grown Avena sativa L. Planta 164: 521–528 (1985).
Vierstra RD, Cordonnier M-M, Pratt LH, Quail PH: Native phytochrome: immunoblot analysis of relative molecular mass and in vitro proteolytic degradation for several plant species. Planta 160: 521–528 (1984).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Heyer, A., Gatz, C. Isolation and characterization of a cDNA-clone coding for potato type A phytochrome. Plant Mol Biol 18, 535–544 (1992). https://doi.org/10.1007/BF00040669
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00040669