Advertisement

Plant Molecular Biology

, Volume 47, Issue 6, pp 805–813 | Cite as

POR C of Arabidopsis thaliana: a third light- and NADPH-dependent protochlorophyllide oxidoreductase that is differentially regulated by light

  • Qingxiang Su
  • Geneviève Frick
  • Gregory Armstrong
  • Klaus Apel
Article

Abstract

During the sequencing of the genome of Arabidopsis thaliana a gene has been identified that encodes a novel NADPH-protochlorophyllide oxidoreductase (POR)-like protein (accession number AC 002560). This protein has been named POR C. We have expressed the POR C protein in Escherichia coli and have determined its in vitro activity. POR C shows the characteristics of a light-dependent and NADPH-requiring POR similar to POR A and POR B. The expression of the POR C gene differs markedly from that of the POR A and POR B genes. In contrast to the POR A and POR B mRNAs, the POR C mRNA has been shown previously to accumulate only after the beginning of illumination. In light-adapted mature plants only POR B and POR C mRNAs were detectable. The amounts of both mRNAs show pronounced diurnal rhythmic fluctuations. While the oscillations of POR B mRNA are under the control of the circadian clock, those of POR C mRNA are not. Another difference between POR B and POR C was found in seedlings that were grown under continuous white light. The concentration of POR C mRNA rapidly declined and soon dropped beyond the limit of detection, after these seedlings were transferred to the dark. On the other hand, POR B mRNA was unaffected by this light/dark shift. When seedlings were exposed to different light intensities, the amounts of POR B mRNA remained the same, while POR A and POR C mRNAs were modulated in an inverse way by these light intensity changes. POR A mRNA was still detectable in seedlings grown under low light intensities but disappeared at higher light intensities, while the mRNA concentration of POR C rose with increasing light intensities. These different responses to light suggest that the functions of the three PORs of Arabidopsis are not completely redundant, but may allow the plant to adapt its needs for chlorophyll biosynthesis more selectively by using preferentially one of the three enzymes under a given light regime.

chlorophyll biosynthesis chloroplast formation NADPH-protochlorophyllide oxidoreductase C 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adamson, H.J., Hiller, R.G. and Walmsley, J. 1997. Protochloro-phyllide reduction and greening in angiosperms: an evolutionary perspective. Photochem. Photobiol. 41: 201–221.Google Scholar
  2. Apel, K. 2001. Chlorophyll biosynthesis. Metabolism and strate-gies of higher plants to avoid photooxidative stress. In: B. Andersson and E.M. Aro (Eds) Regulatory Aspects of Photosyn-thesis, Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 235–252.Google Scholar
  3. Apel, K., Santel, H.J., Redlinger, T.E. and Falk, H. 1980. The protochlorophyllide holochrome of barley (Hordeum vul-gare L.). Isolation and characterization of the NADPH-protochlorophyllide oxidoreductase. Eur. J. Biochem. 111: 251–258.Google Scholar
  4. Armstrong, G.A. 1998. Greening in the dark: light-independent chlorophyll biosynthesis from anoxygenic photosynthetic bac-teria to gymnosperms. J. Photochem. Photobiol. B. Biol. 43: 87–100.Google Scholar
  5. Armstrong, G.A., Runge, S., Frick, G., Sperling, U. and Apel, K. 1995. Identification of NADPH:protochlorophyllide oxidore-ductases A and B: a branched pathway for light-dependent chlorophyll biosynthesis in Arabidopsis thaliana. Plant Physiol. 108: 1505–1517.Google Scholar
  6. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seiod-man, J.G., Smith, J.A. and Struhl, K. 1994. Current Protocols in Molecular Biology. Green/Wiley, New York.Google Scholar
  7. Benli, M., Schulz, R. and Apel, K. 1991. Effect of light on the NADPH-protochlorophyllide oxidoreductase of Arabidopsis thaliana. Plant Mol. Biol. 16: 615–625.Google Scholar
  8. Burke, D.H., Alberti, M. and Hearst, J.E. 1993. bchFNBH bacte-riochlorophyll synthesis genes of Rhodobacter capsulatus and identification of the third subunit of light-independent pro-tochlorophyllide reductase in bacteria and plants. J. Bact. 175: 2414–2422.Google Scholar
  9. Choquet, Y., Rahire, M., Girard-Bascou, J., Erickson, J. and Rochaix, J.-D. 1992. A chloroplast gene is required for the light-independent accumulation of chlorophyll in Chlamydomonas reinhardtii. EMBO J. 11: 1697–1704.Google Scholar
  10. Dehesh, K., Klaas, M., Häuser, I. and Apel, K. 1986. Light-induced changes in the distribution of the 36000-Mr polypeptide of NADPH-protochlorophyllide oxidoreductase within different cellular compartments of barley (Hordeum vulgare L.). I. Lo-calization by immunoblotting in isolated plastids and total leaf extracts. Planta 169: 162–171.Google Scholar
  11. Feierabend, J. and Dehne, S. 1996. Fate of the porphyrin cofactors during light-dependent turnover of catalase and of the photosys-tem II reaction-center protein D1 in mature rye plants. Planta 198: 413–422.Google Scholar
  12. Forreiter, C. and Apel, K. 1993. Light-independent and light-dependent proto-chlorophyllide-reducing activities and two dis-tinct NADPH-protochlorophyllide oxidoreductase polypeptides in mountain pine (Pinus mugo). Planta 190: 536–545.Google Scholar
  13. Forreiter, C., van Cleve, B., Schmidt A. and Apel, K. 1990. Evi-dence for a general light-dependent negative control of NADPH-protochlorophyllide oxidoreductase in angiosperms. Planta 183: 126–132.Google Scholar
  14. Fujita, Y. 1996. Protochlorophyllide reduction: a key step in the greening of plants. Plant Cell Physiol. 37: 411–421.Google Scholar
  15. Fujita, Y., Takagi, H. and Hase, T. 1998. Cloning of the gene en-coding a protochlorophyllide reductase: the physiological signif-icance of the co-existence of light-dependent and-independent protochlorophyllide reduction systems in the cyanobacterium Plectonema boryanum. Plant Cell Physiol. 39: 177–185.Google Scholar
  16. Griffiths, W.T. 1978. Reconstitution of chlorophyllide formation by isolated etioplast membranes. Biochem. J. 174: 681–692.Google Scholar
  17. Hendry, G. and Stobart, A.K. 1986. Chlorophyll turnover in green-ing barley. Phytochemistry 25: 2735–2737.Google Scholar
  18. Holtorf, H., Reinbothe, S., Reinbothe, C., Bereza, B. and Apel, K. 1995. Two routes of chlorophyllide synthesis that are differen-tially regulated by light in barley (Hordeum vulgare L.). Proc. Natl. Acad. Sci. USA 92: 3254–3258.Google Scholar
  19. Ikeuchi, M. and Murakami, S. 1982. Behavior of the 36000-dalton protein in the internal membranes of squash etioplasts during greening. Plant Cell Physiol. 23: 575–583.Google Scholar
  20. Kuroda, H., Masuda, T., Fusada, N. Ohta, H. and Takamiya, K. 1995. Light-enhanced gene expression of NADPH-protochlorophyllide oxidoreductase in cucumber. Biochem. Biophys. Res. Com. 210: 310–316.Google Scholar
  21. Kuroda, H., Masuda, T., Fusada, N. Ohta, H. and Takamiya, K. 2000. Expression of NADPH-protochlorophyllide oxidoreduc-tase in fully green leaves of cucumber. Plant Cell Physiol. 41: 226–229.Google Scholar
  22. Li, J. and Timko, M.P. 1996. The pc-1 phenotype of Chlamy-domonas reinhardtii results from a deletion in the nuclear gene for NADPH:protochlorophyllide oxidoreductase. Plant Mol. Biol. 30: 15–37.Google Scholar
  23. Li, J., Goldschmidt-Clermont, M. and Timko, M.P. 1993. Chloroplast-encoded chlB is required for light-independent pro-tochlorophyllide reductase activity in Chlamydomonas rein-hardtii. Plant Cell 5: 1817–1829.Google Scholar
  24. Lindholm, J. and Gustafsson, P. 1991. Homologues of the green algal gidA gene and the liverwort frxC gene are present in the chloroplast genomes of conifers. Plant Mol. Biol. 17: 787–798.Google Scholar
  25. Mapleston, E.R. and Griffiths, W.T. 1980. Light modulation of the activity of the protochlorophyllide oxidoreductase. Biochem. J. 189: 125–133.Google Scholar
  26. Oosawa, N., Masuda, T., Awai, K., Fusada, N., Shimada, H., Ohta, H. and Takamiya, K. 2000. Identification and light-induced expression of a novel gene of NADPH-protochlorophyllide oxi-doreductase isoform in Arabidopsis thaliana. FEBS Lett. 474: 133–136.Google Scholar
  27. Reinbothe, S., Reinbothe, C., Lebedev, N. and Apel, K. 1996. POR A and POR B, two light-dependent protochlorophyllide-reducing enzymes of angiosperm chlorophyll biosynthesis. Plant Cell 8: 763–769.Google Scholar
  28. Roitgrund, C. and Mets, L. 1990. Localization of two novel chloro-plast functions: trans-splicing of RNA and protochlorophyllide reduction. Curr. Genet. 17: 147–153.Google Scholar
  29. Runge, S., Sperling, U., Frick, G., Apel, K. and Arm-strong, G.A. 1996. Distinct roles for light-dependent NADPH-protochlorophyllide oxidoreductases (POR) A and B during greening in higher plants. Plant J. 9: 513–523.Google Scholar
  30. Santel, H.J. and Apel, K. 1981. The protochlorophyllide holochrome of barley (Hordeum vulgare L.). The effect of light on the NADPH-protochlorophyllide oxidoreductase. Eur. J. Biochem. 120: 95–103.Google Scholar
  31. Skinner, J.S. and Timko, M.P. 1999. Differential expression of genes encoding the light-dependent and light-independent enzymes for protochlorophyllide reduction during development in loblolly pine. Plant Mol. Biol. 39: 577–592.Google Scholar
  32. Spano, A.J., He Z.-H., Michel, H., Hunt, D.F. and Timko, M.P. 1992a. Molecular cloning, nuclear gene structure, and develop-mental expression of NADPH:protochlorophyllide oxidorectase in pea (Pisum sativum L.). Plant Mol. Biol. 18: 967–972.Google Scholar
  33. Spano, A.J., He Z.-H. and Timko, M.P. 1992 b. NADPH:protochlorophyllide oxidoreductases in white pine (Pinus strobus) and loblolly pine (P. taeda). Mol. Gen. Genet. 236: 86–95.Google Scholar
  34. Sperling, U., van Cleve, B., Frick, G., Apel, K. and Armstron G.A. 1997. Overexpression of light-dependent POR A or POR B in plants depleted of endogenous POR by far-red light en-hances seedling survival in white light and protects against photooxidative damage. Plant J. 12: 649–658.Google Scholar
  35. Sperling, U., Franck, F., van Cleve, B., Frick, G., Apel, K. and Arm-strong, G.A. 1998. Etioplast differentiation in Arabidopsis: both.813 POR A and POR B restore the prolamellar body and photoac-tive protochlorophyllide-F655 to the cop1 photomorphogenic mutant. Plant Cell 10: 283–296.Google Scholar
  36. Suzuki, J.Y. and Bauer, C.E. 1995. A prokaryotic origin for light-dependent chlorophyll biosynthesis of plants. Proc. Natl. Acad. Sci. USA 92: 3749–3753.Google Scholar
  37. Suzuki, J.Y. and Bauer, C.E. 1992. Light-independent chlorophyll biosynthesis: involvement of the chloroplast gene chlL (frxC). Plant Cell 4: 929–940.Google Scholar
  38. Takio, S., Nakao, N., Suzuki, T., Tanaka, K., Yamamoto, I. and Satoh, T. 1998. Light-dependent expression of protochlorophyl-lide oxidoreductase gene in the liverwort, Marchantia paleacea var. diptera. Plant Cell Physiol. 39: 665–669.Google Scholar
  39. Thomas, H. 1997. Chlorophyll: a symptom and a regulator of plastid development. New Phytol. 136: 163–181.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Qingxiang Su
    • 1
  • Geneviève Frick
    • 1
  • Gregory Armstrong
    • 1
  • Klaus Apel
    • 1
  1. 1.Institute of Plant Sciences, Swiss Federal Institute of Technology (ETH)ZürichSwitzerland

Personalised recommendations