Abstract
Overexpression of phytochrome A results in an increased inhibition of hypocotyl elongation under red and far-red light. We used this approach to assay for the function of N-terminal mutations of rice (Oryza sativa L.) phytochrome A. Transgenic tobacco seedlings that express the wild-type rice phytochrome A (RW), a rice phytochrome A lacking the first 80 amino acids (NTD) or a rice phytochrome A with a conversion of the first 10 serines into alanine residues (S/A) were compared with untransformed wild-type tobacco (Nicotiana tabacum L. cv. Xanthi) seedlings. Experiments under different fluence rates showed that RW and, even more strongly, S/A increased the response under both red and far-red light, whereas NTD decreased the response under far-red light but hardly altered the response under red light. These results indicate that NTD not only lacks residues essential for an increased response under red light but also distorts the wild-type response under far-red light. Wild-type rice phytochrome A and, even more so, S/A mediate an enhanced phytochrome A as well as phytochrome B function, whereas NTD interferes with the function of endogenous tobacco phytochrome A as well as that of rice phytochrome A when co-expressed in a single host. Experiments with seedlings of different ages and various times of irradiation under far-red light demonstrated that the effect of NTD is dependent on the stage of development. Our results suggest that the lack of the first 80 amino acids still allows a rice phytochrome A to interact with the phytochrome transduction pathway, albeit nonproductively in tobacco seedlings.
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Abbreviations
- HIR:
-
high-irradiance response
- NTD:
-
N-terminal deletion mutant of rice phytochrome A
- Pfr:
-
far-red-absorbing form of phytochrome
- Pr:
-
red-absorbing form of phytochrome
- RW:
-
rice wild-type phytochrome A
- S/A:
-
serine-to-alanine mu-tant of rice phytochrome A
- wNTD:
-
weakly expressing NTD line
- XAN:
-
wild-type tobacco cv. Xanthi
References
Boylan, M.T., Quail, P.H. (1989) Oat phytochrome is biologically active in transgenic tomatoes. Plant Cell 1, 765–773
Boylan, M.T., Quail, P.H. (1991) Phytochrome A overexpression inhibits hypocotyl elongation in transgenic Arabidopsis. Proc. Natl. Acad. Sci. USA 88, 10806–10810
Boylan, M.T., Douglas, N., Quail, P.H. (1994) Dominant negative suppression of Arabidopsis photoresponses by mutant phytochrome A sequences identifies spatially discrete regulatory domains in the photoreceptor. Plant Cell 6, 449–460
Chai, Y.-G., Song, P.-S., Cordonnier, M.M., Pratt, L.H. (1987) A photoreversible circular dichroism spectral change in oat phytochrome is suppressed by a monoclonal antibody that binds near its N-terminus and by chromophore modification. Biochemistry 26, 4947–4952
Cherry, J.R., Hershey, H.P., Vierstra, R.D. (1991) Characterization of tobacco expressing functional oat phytochrome. Plant Physiol. 96, 775–785
Cherry, J.R., Hondred, D., Walker, J.M., Vierstra, R.D. (1992) Phytochrome requires the 6-kDa N-terminal domain for full biological activity. Proc. Natl. Acad. Sci. USA 89, 5039–5043
Cordonnier, M.M., Greppin, H., Pratt, L.H. (1985) Monoclonal antibodies with different affinities to the red-absorbing and far-red-absorbing forms of phytochrome. Biochemistry 24, 3246–3253
Frohnmeyer, H., Ehmann, B., Kretsch, T., Rocholl, M., Harter, K., Nagatani, A., Furuya, M., Batschauer, A., Hahlbrock, K., Schäfer, E. (1992) Differential usage of photoreceptors for chalcone synthase gene expression during plant development. Plant J. 2, 899–906
Furuya, M., ed. (1987) Phytochrome and photoregulation in plants. Academic Press, Tokyo
Furuya, M. (1989) Molecular properties and biogenesis of phytochrome I and II. Arch. Biophys, 25, 133–137
Furuya, M. (1993) Phytochromes: Their molecular species, gene families and function. Annu. Rev. Plant Physiol. Plant Mol. Biol. 44, 617–645
Grimm, R., Eckerskorn, C., Lottspeich, F., Zenger, C., Rüdiger, W. (1988) Sequence analysis of proteolytic fragments of 124-ilodalton phytochrome from etiolated Avena sativa L.: Conclusions on the conformations of the native protein. Planta 174, 396–401
Hartmann, K.M. (1966) A general hypothesis to interpret the high energy phenomena of photomorphogenesis on the basis of phytochrome. Photochem. Photobiol. 5, 349–366
Herskowitz, I. (1987) Functional inactivation of genes by dominant negative mutations. Nature 329, 219–222
Jones, A.M., Vierstra, R.D., Daniels, S.M., Quail, P.H. (1985) The role of separate molecular domains in the structure of phytochrome from etiolated Avena sativa L. Planta 164, 501–506
Kay, S.A., Nagatani, A., Keith, B., Deak, M., Furuya, M., Chua, N.-H. (1989) Rice phytochrome is biologically active in transgenic tobacco. Plant Cell 1, 775–782
Keller, J.M., Shanklin, J., Vierstra, R.D., Hershey, H.P. (1989) Expression of a functional monocotyledonous phytochrome in transgenic tobacco. EMBO J. 8, 1005–1012
Kendrick, R.E., Kronenberg, G.H.M., eds. (1994) Photomorphogenesis in Plants. Martinus Nijhoff Publishers Dordrecht, The Netherlands
Kunkel, T., Tomizawa, K.-I., Kern, R., Furuya, M., Chua, N.-H., Schäfer, E. (1993) In vitro formation of a photoreversible adduct of phycocyanobilin and tobacco apophytochrome B. Eur. J. of Biochem. 215, 587–594
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
Lagarias, J.C., Lagarias, D.M. (1989) Self-assembly of synthetic phytochrome holoprotein in vitro. Proc. Natl. Acad. Sci. USA 86, 5778–5780
Lagarias, J.C., Mercurio, F.M. (1985) Structure and function studies on phytochrome. J. Biol. Chem. 260, 2415–2423
McCormac, A.C., Cherry, J.R., Hershey, H.P., Vierstra, R.D., Smith, H. (1991) Photoresponses of transgenic tobacco plants expressing an oat phytochrome gene. Planta 185, 162–170
McCormac, A.C., Whitelam, G.C., Smith, H. (1992) Light grown plants of transgenic tobacco expressing an oat phytochrome A gene under the control of a constitutive viral promotor exhibit persistent growth inhibition by far-red light. Planta 188, 173–181
McCormac, A.C., Wagner, D., Boylan, M.T., Quail, P.H., Smith, H., Whitelam, G.C. (1993) Photoresponses of transgenic Arabidopsis seedlings expressing introduced phytochrome B-encoding cDNAs: evidence that phytochrome A and phytochrome B have distinct photoregulatory functions. Plant J. 4, 19–27
Nagatani, A., Yamamoto, K., Furuya, M., Fukumoto, T., Yamashita, A. (1984) Production and characterization of monoclonal antibodies which distinguish different surface structures of pea phytochrome. Plant Cell Physiol. 25, 1059–1068
Nagatani, A., Kay, S.A., Deak, M., Chua, N.-H., Furuya, M. (1991) Rice type I phytochrome regulates hypocotyl elongation in transgenic tobacco seedlings. Proc. Natl. Acad. Sci. USA 88, 5207–5211
Nagatani, A., Reed, J.W., Chory, J. (1993a) Isolation and initial characterization of Arabidopsis mutants that are deficient in phytochrome A. Plant Physiol. 102, 269–277
Nagatani, A., Nishizawa, N.K., Mori, S., Kay, S.A., Chua, N.-H., Furuya, M. (1993b) Light regulation of hypocotyl elongation and greening in transgenic tobacco seedlings that over-express rice phytochrome A. Plant Cell Physiol. 34, 825–833
Parks, B., Quail, P.H. (1993) hy 8, a new class of Arabidopsis long hypocotyl mutants deficient in functional phytochrome A. Plant Cell 5, 39–48
Pratt, H., Marmé, D. (1976) Red light-enhanced phytochrome pelletability. Re-examination and further characterization. Plant Physiol. 58, 686–692
Quail, P.H. (1991) Phytochrome: a light-activated molecular switch that regulates plant gene expression. Annu. Rev. Genet 25, 389–409
Reed, J.W., Nagpal, P., Poole, D.S., Furuya, M., Chory, J. (1993) Mutations in the gene for the red/far-red light receptor phytochrome B alter cell elongation and physiological responses throughout Arabidopsis development. Plant Cell 5, 147–157
Schäfer, E. (1975) A new approach to explain the “high irradiance response” of photomorphogenesis on the basis of phytochrome. J. Math. Biol. 2, 41–56
Schäfer, E. (1977) Kunstlicht und Pflanzenzucht. In: Optische Strahlungsquellen, pp. 249–266, Albrecht, H., ed. Lexika-Verlag, Grafenau, Germany
Sharrock, R.A., Quail, P.H. (1989) Novel phytochrome sequences in Arabidopsis thaliana: structure, evolutition and differential expression of a plant regulatory photoreceptor family. Genes Dev. 3, 1745–1757
Stockhaus, J, Nagatani, A., Halfter, U., Kay, S.A., Furuya, M., Chua, N.-H. (1992) Serine-to-alanine substitutions at the aminoterminal region of phytochrome A result in an increase in biological activity. Gen. Dev. 6, 2364–2372
Vierstra, R.D., Quail, P.H. (1986), The protein In: Photomorphogenesis in plants, pp. 35–59, Kendrick, R.E., Kronenberg G.H.M., eds. Nijhoff, Amsterdam
Vierstra, R.D., Quail, P.H., Hahn, T.-R., Song, P.-S. (1987) Comparison of the protein conformations between different forms (Pr and Pfr) of native (124 kD) and degraded (118/114 kD) phytochrome from Avena sativa L. Photochem. Photobiol. 45, 429–432
Whitelam, G.C., Johnson, E., Peng, J., Carol, P., Anderson, M.L., Cowl, J.S., Harberd, N.P. (1993) Phytochrome A null mutants of Arabidopsis display a wild-type phenotype in white light. Plant Cell 5, 757–768
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We thank Masaki Furuya (Adv. Research Laboratory, Hitachi, Saitama, Japan) and Akira Nagatani (RIKEN Institute, Saitama, Japan) for providing the monoclonal antibodies mAP5 and mAR14. The work was supported by a grant from the Human Frontier Science Program. K.E. was a recipient of a “Landesgraduiertenförderung” fellowship.
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Emmler, K., Stockhaus, J., Chua, NH. et al. An amino-terminal deletion of rice phytochrome A results in a dominant negative suppression of tobacco phytochrome A activity in transgenic tobacco seedlings. Planta 197, 103–110 (1995). https://doi.org/10.1007/BF00239945
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DOI: https://doi.org/10.1007/BF00239945