Skip to main content
Log in

NADPH : protochlorophyllide oxidoreductases in white pine (Pines strobes) and loblolly pine (P. taeda)

Evidence for light and developmental regulation of expression and conservation in gene organization and protein structure between angiosperms and gymnosperms

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

NADPH : protochlorophyllide oxidoreductase (pchlide reductase, EC 1.6.99.1) catalyzes the light-dependent reduction of protochlorophyllide in higher plants. Cloned cDNAs encoding two distinct pchlide reductases were isolated from a λgt11 library constructed from poly(A)+ RNA prepared from the cotyledons of dark-grown white pine (Pines strobes) seedlings and a nuclear gene (lpcr) analogous to one of these cDNAs has been characterized from loblolly pine (P. taeda). The pine gene encodes an approximately 43 kDa precursor polypeptide consisting of a 334-amino acid mature protein and a 66-amino acid transit peptide. The deduced primary structures for the pine proteins are highly homologous to those reported from monocots and dicots. The coding portion of the pine lpcr gene is interrupted by four introns. The placement of these introns within the pine lpcr gene is identical to that observed in pea (Pisum sativum), suggesting conservation in gene organization between dicot and gymnosperm species. Western blot analysis using polyclonal antiserum against oat pchlide reductase detected in extracts of dark-grown pine cotyledons a single immunoreactive protein, which declined in abundance during a 48 h period of illumination with white light. Cotyledons of dark-grown seedlings were also found to accumulate high levels of pchlide reductase mRNA; however, little or no change in the steady-state levels of mRNA encoding pchlide reductase was observed in these tissues following illumination. Stem tissue of dark-grown seedlings did not contain significant levels of pchlide reductase mRNA, whereas stems of light-grown plants of the same age accumulated substantial amounts of the message. These results suggest that light and the developmental age of the tissue affect regulation of lpcr expression in pine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alosi MC, Neale DB, Kinlaw CS (1990) Expression of cab genes in Douglas-fir is not strongly regulated by light. Plant Physiol 93:829–833

    Google Scholar 

  • Apel K (1981) The protochlorophyllide holochrome of barley (Hordeum vulgare L.). Phytochrome-induced decrease of translatable mRNA coding for the NADPH-protochlorophyllide oxidoreductase. Eur J Biochem 120:89–93

    Google Scholar 

  • Apel K, Sanel J-J, Redlinger TE, Falk H (1980) The protochlorophyllide holochrome of barley (Hordeum vulgare L.). Isolation and characterization of the NADPH protochlorophyllide oxidoreductase. Eur J Biochem 111:251–258

    Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 60:1–15

    Google Scholar 

  • Batschauer A, Apel K (1984) An inverse control by phytochrome of the expression of two nuclear genes in barley (Hordeum vulgare L.). Eur J Biochem 143:593–597

    Google Scholar 

  • Benli M, Schulz R, Apel K (1991) Effect of light on the NADPH-protochlorophyllide oxidoreductase of Arabidopsis thaliana. Plant Mol Biol 16:615–625

    Google Scholar 

  • Boese QF, Spano AJ, Li J, Timko MP (1991) Amino levulinic acid dehydratase in pea (Pisum sativum L.). Identification of an unusual metal-binding domain in the plant enzyme. J Biol Chem 266:17060–17066

    Google Scholar 

  • Bogdanovic M (1973) Chlorophyll formation in the dark. 1. Chlorophyll in pine seedlings. Physiol Plant 29:17–18

    Google Scholar 

  • Bogorad L (1950) Factors associated with the synthesis of chlorophyll in the dark in seedlings of Pinus jeffreyi. Bot Gaz 111:221–241

    Google Scholar 

  • Brown JWS (1986) A catalogue of splice junction and putative branch point sequences from plant introns. Nucleic Acids Res 14:9549–9559

    Google Scholar 

  • Cashmere AR (1982) The isolation of polyA+ messenger RNA from higher plants. In: Edelman M, Hallick RB, Chua N-H (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, Amsterdam, pp 387–392

    Google Scholar 

  • Darrah PM, Kay SA, Teakle GR, Griffiths WT (1990) Cloning and sequencing of protochlorophyllide reductase. Biochem J 265:789–798

    Google Scholar 

  • Dehesh K, Klaas M, Hauser I, Apel K (1986a) Light-induced changes in the distribution of the 36000-Mr polypeptide of the NADPH-protochlorophyllide oxidoreductase within different cellular compartments of barley (Hordeum vulgare L.). I. Localization by immunoblotting in isolated plastids and total leaf extracts. Planta 169:162–171

    Google Scholar 

  • Dehesh K, von Cleve B, Ryberg M, Apel K (1986b) Light-induced changes in the distribution of the 36000-Mr polypeptide of the NADPH-protochlorophyllide oxidoreductase within different cellular compartments of barley (Hordeum vulgare L.). II. Localization by immunogold labelling in ultrathin sections. Planta 169:172–183

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    Google Scholar 

  • Ford C, Wang W-Y (1980) Three new yellow loci in Chlamydomonas reinhardtii. Mol Gen Genet 179:259–263

    Google Scholar 

  • Forreiter C, van Cleve B, Schmidt A, Apel K (1991) Evidence for a general mechanism of light-dependent control of the NADPH-protochlorophyllide oxidoreductase in higher plants. Planta 183:126–132

    Google Scholar 

  • Girard-Bascou J (1987) Mutation in four chloroplast loci of Chlamydomonas reinhardtii affecting photosystem I reaction centers. Curr Genet 12:483–488

    Google Scholar 

  • Griffiths WT (1974) Protochlorophyll and protochlorophyllide as precursors for chlorophyll synthesis in vitro. FEBS Lett 49:196–200

    Google Scholar 

  • Griffiths WT (1978) Reconstitution of chlorophyllide formation by isolated etioplast membranes. Biochem J 174:681–692

    Google Scholar 

  • Griffiths WT, Walker CJ (1987) In: BigginsJ (ed) Progress in photosynthesis research. Balban International Science Services, Philadelphia, pp 65–71

    Google Scholar 

  • Gubler U, Hoffman BJ (1983) A simple and very efficient method for generating cDNA libraries. Gene 25:263–269

    Google Scholar 

  • Harpster M, Apel K (1985) The light-dependent regulation of gene expression during plastid development in higher plants. Physiol Plant 64:147–152

    Google Scholar 

  • Hauser I, Dehesh K, Apel K (1984) The proteolytic degradation in vitro of the NADPH-protochlorophyllide oxidoreductase of barley (Hordeum vulgare L.). Arch Biochem Biophys 228:557–586

    Google Scholar 

  • Henikoff S (1987) Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol 155:156–165

    Google Scholar 

  • Ikeuchi M, Murakami S (1982) Behavior of the 36 000 dalton protein in the internal membranes of squash etioplasts during greening. Plant Cell Physiol 23:575–583

    Google Scholar 

  • Jelic G, Bogdanovic M (1988) Antagonism between abscisic acid and cytokinin in chlorophyll synthesis in pine. Plant Sci 61:197–202

    Google Scholar 

  • Joyard J, Block M, Pineau B, Albrieux C, Douce R (1990) Envelope membranes from mature spinach chloroplasts contain a NADPH: protochlorophyllide reductase on the cytosolic side of the outer membrane. J Biol Chem 265:21820–21827

    Google Scholar 

  • Kay SA, Griffiths WT (1983) Light-induced breakdown of NADPH-protochlorophyllide oxidoreductase in vitro. Plant Physiol 72:229–236

    Google Scholar 

  • Kittsteiner U, Paulsen H, Schendel R, Rudiger W (1990) Lack of light regulation of NADPH: protochlorophyllide oxido-reductase mRNA in cress seedlings (Lepidium sativum L.). Z Naturforsch 45c:1077–1079

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Lidholm J, Gustafsson P (1991) Homologues of the green algal gidA gene and the liverwort frxC gene are present on the chloroplast genomes of conifers. Plant Mol Biol 17:787–798

    Google Scholar 

  • Mapleston ER, Griffiths WT (1980) Light modulation of the activity of protochlorophyllide reductase. Biochem J 189:125–133

    Google Scholar 

  • Meyer G, Bliedung H, Kloppstech K (1983) NADPH-protochlorophyllide oxidoreductase: reciprocal regulation in mono- and dicotyledonean plants. Plant Cell Reports 2:26–29

    Google Scholar 

  • Moller W, Anions R (1985) Phosphate-binding sequences in nucleotide binding proteins. FEBS Lett 186:1–7

    Google Scholar 

  • Mosinger E, Batschauer A, Schafer E, Apel K (1985) Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare). Eur J Biochem 147:137–142

    Google Scholar 

  • Mukai Y, Yamamoto N, Koshiba T (1991) Light-independent and tissue-specific accumulation of light-harvesting chlorophyll a/b binding protein and ribulose bisphosphate carboxylase in darkgrown pine seedlings. Plant Cell Physiol 32:1303–1306

    Google Scholar 

  • Niklolic D, Bogdanovic M (1972) Plastid differentiation and chlorophyll synthesis in cotyledons of black pine seedlings grown in the dark. Protoplasma 75:205–213

    Google Scholar 

  • Oliver RP, Griffiths WT (1981) Covalent labelling of the NADPH-protochlorophyllide oxidoreductase from etioplast membranes with (3H)N-phenylmaleimide. Biochem J 195:93–101

    Google Scholar 

  • On K, Packer N, Adamson H (1990) Immunodetection and photostability of NADPH-protochlorophyllide oxidoreductase in Pinus pinea L. Photosynthesis Res 23:89–94

    Google Scholar 

  • Parthier B, Lerbs S, Klyachko NL (1981) Plastogenesis and cytokinin action. In: Guern J, Peaud-Lenoel C (eds) Metabolism and molecular activity of cytokinins. Springer-Verlag, New York, pp 275–286

    Google Scholar 

  • Perlman D, Halvorson HO (1983) A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. J Mol Biol 167:391–409

    Google Scholar 

  • Roitgrund C, Mets LJ (1990) Localization of two novel chloroplast genome functions: trans-splicing of RNA and protochlorophyllide reduction. Curr Genet 17:147–153

    Google Scholar 

  • Sager R (1961) Photosynthetic pigments in mutant strains of Chlamydomonas. Carnegie Inst Wash Yearbook 60:374–376

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (eds) (1989) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Santel H-J, 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 

  • Schulz R, Steinmuller K, Klaas M, Forreiter C, Rasmussen S, Hiller C, Apel K (1989) Nucleotide sequence of a cDNA coding for the NADPH-protochlorophyllide oxidoreductase (PCR) of barley (Hordeum vulgare L.) and its expression in Escherichia coli. Mol Gen Genet 217:355–361

    Google Scholar 

  • Selstam E, Widell A, Johannsan B-A (1987) A comparison of prolamellar bodies for wheat, Scots pine and Jeffrey pine. Pigment spectra and properties of protochlorophyllide oxidoreductase. Physiol Plant 70:209–214

    Google Scholar 

  • Shaw P, Henwood J, Oliver R, Griffiths T (1985) Immunogold localisation of protochlorophyllide oxidoreductase in barley etioplasts. Eur J Cell Biol 39:50–55

    Google Scholar 

  • Spano AJ, Timko MP (1991) Isolation, characterization, and partial amino acid sequence of a chloroplast-localized porphobilinogen deaminase from pea (Pisum sativum L.). Biochem Biophys Acta 1076:29–36

    Google Scholar 

  • Spano AJ, He Z, Michel H, Hunt DF, Timko MP (1992) Molecular cloning, nuclear gene structure, and developmental expression of NADPH-protochlorophyllide oxidoreductase in pea (Pisum sativum L.). Plant Mol Biol 18:967–972

    Google Scholar 

  • Yamamoto N, Mukai Y, Matsuoka M, Kano-Murakami Y, Tanaka Y, Ohashi Y, Odani K (1991) Light-independent expression of cab and rbcS genes in dark-grown pine seedlings. Plant Physiol 95:379–383

    Google Scholar 

  • Zurawski G, Perrot B, Bottomley W, Whitfeld PR (1981) The structure of the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase from spinach chloroplast DNA. Nucleic Acids Res 9:3251–3270

    Google Scholar 

  • Zurawski G, Bohnert H, Whitfeld PR, Bottomley W (1982) Nucleotide sequence of the gene for the Mr 32 000 thylakoid membrane protein from Spinach oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of Mr 38 950. Proc Natl Acad Sci USA 79:7699–7703

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by R.G. Herrmann

Rights and permissions

Reprints and permissions

About this article

Cite this article

Spano, A.J., He, Z. & Timko, M.P. NADPH : protochlorophyllide oxidoreductases in white pine (Pines strobes) and loblolly pine (P. taeda). Molec. Gen. Genet. 236, 86–95 (1992). https://doi.org/10.1007/BF00279646

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00279646

Key words

Navigation