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. 1995 Sep;109(1):231–238. doi: 10.1104/pp.109.1.231

Correlated Changes in the Activity, Amount of Protein, and Abundance of Transcript of NADPH:Protochlorophyllide Oxidoreductase and Chlorophyll Accumulation during Greening of Cucumber Cotyledons.

K Yoshida 1, R M Chen 1, A Tanaka 1, H Teramoto 1, R Tanaka 1, M P Timko 1, H Tsuji 1
PMCID: PMC157580  PMID: 12228591

Abstract

Changes in the activity and abundance of NADPH:protochlorophyllide oxidoreductase (NPR) and the abundance of mRNA encoding it were examined during the greening of 5-d-old etiolated cucumber cotyledons under continuous illumination. To measure NPR activity in the extracts from fully greened tissues, we have developed an improved method of assay. Upon exposure of etiolated cotyledons to light, NPR activity decreased rapidly within the first 2 h of exposure. Thereafter, enzymatic activity increased transiently, reaching a submaximum level at 12 h, and decreased slowly. The level of immunodetectable NPR protein followed the same pattern of changes during 96 h of greening as observed for NPR activity. The NPR mRNA in etiolated cotyledons disappeared quickly in the 1st h of irradiation. However, the level of mRNA increased thereafter to reach 3-fold or more of the dark level at 12 h and then decreased. The changes in the activity, protein level, and mRNA level after the first rapid decreases corresponded chronologically and nearly paralleled the increase in the rate of chlorophyll accumulation. These findings suggest that the greening of cucumber cotyledons is regulated basically by the level of NPR protein without activation or repression of enzymatic activity and that NPR mRNA increased by light maintains the level of enzyme protein necessary for greening.

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Selected References

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  1. Apel K., Santel H. J., Redlinger T. E., Falk H. The protochlorophyllide holochrome of barley (Hordeum vulgare L.). Isolation and characterization of the NADPH:protochlorophyllide oxidoreductase. Eur J Biochem. 1980 Oct;111(1):251–258. doi: 10.1111/j.1432-1033.1980.tb06100.x. [DOI] [PubMed] [Google Scholar]
  2. Apel K. The protochlorophyllide holochrome of barley (Hordeum vulgare L.). Phytochrome-induced decrease of translatable mRNA coding for the NADPH: protochlorophyllide oxidoreductase. Eur J Biochem. 1981 Nov;120(1):89–93. doi: 10.1111/j.1432-1033.1981.tb05673.x. [DOI] [PubMed] [Google Scholar]
  3. Batschauer A., Apel K. An inverse control by phytochrome of the expression of two nuclear genes in barley (Hordeum vulgare L.). Eur J Biochem. 1984 Sep 17;143(3):593–597. doi: 10.1111/j.1432-1033.1984.tb08411.x. [DOI] [PubMed] [Google Scholar]
  4. Beer N. S., Griffiths W. T. Purification of the enzyme NADPH: protochlorophyllide oxidoreductase. Biochem J. 1981 Apr 1;195(1):83–92. doi: 10.1042/bj1950083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Benli M., Schulz R., Apel K. Effect of light on the NADPH-protochlorophyllide oxidoreductase of Arabidopsis thaliana. Plant Mol Biol. 1991 Apr;16(4):615–625. doi: 10.1007/BF00023426. [DOI] [PubMed] [Google Scholar]
  6. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  7. Forreiter C., Apel K. Light-independent and light-dependent protochlorophyllide-reducing activities and two distinct NADPH-protochlorophyllide oxidoreductase polypeptides in mountain pine (Pinus mugo). Planta. 1993;190(4):536–545. doi: 10.1007/BF00224793. [DOI] [PubMed] [Google Scholar]
  8. Franck F., Strzalka K. Detection of the photoactive protochlorophyllide-protein complex in the light during the greening of barley. FEBS Lett. 1992 Aug 31;309(1):73–77. doi: 10.1016/0014-5793(92)80742-y. [DOI] [PubMed] [Google Scholar]
  9. Griffiths W. T. Reconstitution of chlorophyllide formation by isolated etioplast membranes. Biochem J. 1978 Sep 15;174(3):681–692. doi: 10.1042/bj1740681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. He Z. H., Li J., Sundqvist C., Timko M. P. Leaf Developmental Age Controls Expression of Genes Encoding Enzymes of Chlorophyll and Heme Biosynthesis in Pea (Pisum sativum L.). Plant Physiol. 1994 Oct;106(2):537–546. doi: 10.1104/pp.106.2.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Holtorf H., Reinbothe S., Reinbothe C., Bereza B., Apel K. Two routes of chlorophyllide synthesis that are differentially regulated by light in barley (Hordeum vulgare L.). Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3254–3258. doi: 10.1073/pnas.92.8.3254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Häuser I., Dehesh K., Apel K. The proteolytic degradation in vitro of the NADPH-protochlorophyllide oxidoreductase of barley (Hordeum vulgare L.). Arch Biochem Biophys. 1984 Feb 1;228(2):577–586. doi: 10.1016/0003-9861(84)90025-0. [DOI] [PubMed] [Google Scholar]
  13. Joyard J., Block M., Pineau B., Albrieux C., Douce R. Envelope membranes from mature spinach chloroplasts contain a NADPH:protochlorophyllide reductase on the cytosolic side of the outer membrane. J Biol Chem. 1990 Dec 15;265(35):21820–21827. [PubMed] [Google Scholar]
  14. Mapleston R. E., Griffiths W. T. Light modulation of the activity of protochlorophyllide reductase. Biochem J. 1980 Jul 1;189(1):125–133. doi: 10.1042/bj1890125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mösinger E., Batschauer A., Schäfer E., Apel K. Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare). Eur J Biochem. 1985 Feb 15;147(1):137–142. doi: 10.1111/j.1432-1033.1985.tb08729.x. [DOI] [PubMed] [Google Scholar]
  16. Pineau B., Gerard-Hirne C., Douce R., Joyard J. Identification of the Main Species of Tetrapyrrolic Pigments in Envelope Membranes from Spinach Chloroplasts. Plant Physiol. 1993 Jul;102(3):821–828. doi: 10.1104/pp.102.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Teakle G. R., Griffiths W. T. Cloning, characterization and import studies on protochlorophyllide reductase from wheat (Triticum aestivum). Biochem J. 1993 Nov 15;296(Pt 1):225–230. doi: 10.1042/bj2960225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yoshida K., Nagano Y., Murai N., Sasaki Y. Phytochrome-regulated expression of the genes encoding the small GTP-binding proteins in peas. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6636–6640. doi: 10.1073/pnas.90.14.6636. [DOI] [PMC free article] [PubMed] [Google Scholar]

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