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Differential distribution of chlorophyll biosynthetic intermediates in stroma, envelope and thylakoid membranes in Beta vulgaris

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Abstract

Stroma, envelope and thylakoid membranes were prepared from chloroplasts isolated from leaves of Beta vulgaris. Out of total plastidic protochlorophyllide, envelope membranes contained 1.5%, thylakoids had the maximum 98.48% and stroma had a trace fraction of 0.02%. Distribution of the Mg-protoporphyrin IX and its monoester was 89.0% in thylakoids, 10.0% in stroma and 1.0% in envelope. A substantial fraction (33.77%) of plastidic protoporphyrin IX was partitioned into stroma. Envelope contained 0.66% and thylakoids had 65.57% of the total plastidic protoporphyrin IX pool. The proportion of monovinyl and divinyl forms of protochlorophyllide was almost similar in intact plastid, thylakoids, and outer and inner envelope membranes suggesting a tight regulation of vinyl reductase enzyme. The significance of differential distribution of chlorophyll biosynthetic intermediates among thylakoids, envelope and stroma is discussed.

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Abbreviations

ALA:

5-Aminolevulinic acid

Chl:

Chlorophyll

Chlide:

Chlorophyllide

DV Pchlide:

Divinyl protochlorophyllide

HEAR:

Hexane extracted acetone residue solvent mixture

LHCP II:

Light harvesting chlorophyll protein complex II

MP(E):

Mg-protoporphyrin IX + its monoester

MV Pchlide:

Monovinyl protochlorophyllide

Pchlide :

Protochlorophyllide

Proto IX:

Protoporphyrin IX

References

  • Armstrong GA, Runge S, Frick G, Sperling U, Apel K (1995) Identification of NADPH:protochlorophyllide oxidoreductases A and B: a branched pathway for light-dependent chlorophyll biosynthesis in Arabidopsis thaliana. Plant Physiol 108:1505–1517

    Article  PubMed  CAS  Google Scholar 

  • Barthelemy X, Bouvier G, Radunz A, Docquier S, Schimd GH, Franck F (2000) Localization of NADPH-protochlorophyllide reductase in plastids of barley at different greening stages. Photosynth Res 64:63–76

    Article  PubMed  CAS  Google Scholar 

  • Block MA, Tewari AK, Albrieux C, Marechal E, Joyard J (2002) The plant S-adenosyl-l-methionine:Mg-protoporphyrin IX methyltransferase is located in both envelope and thylakoid chloroplast membranes. Eur J Biochem 269:240–248

    Article  PubMed  CAS  Google Scholar 

  • Chakraborty N, Tripathy BC (1992) Involvement of singlet oxygen in 5-aminolevulinicacid-induced photodynamic damage of cucumber (Cucumis sativus L.) chloroplasts. Plant Physiol 98:7–11

    PubMed  CAS  Google Scholar 

  • Coves J, Block MA, Joyard J, Douce R (1986) Solubilization and partial purification of UDPgalactose: diacylglycerol galactosyltransferase activity from spinach chloroplast envelope. FEBS Lett 208:401–406

    Article  CAS  Google Scholar 

  • Douce R, Joyard J (1990) Biochemistry and function of the plastid envelope. Annu Rev Cell Biol 6:173–216

    Article  PubMed  CAS  Google Scholar 

  • Eichacker LA, Soll J, Lauterbach P, Rudiger W, Klein RR, Mullet JE (1990) In vitro synthesis of chlorophyll a in the dark triggers accumulation of chlorophyll a apoproteins in barley etioplasts. J Biol Chem 265:13566–13571

    PubMed  CAS  Google Scholar 

  • Grafe S, Saluz HP, Grimm B, Hanel F (1999) Mg-chelatase of tobacco: the role of the subunit CHL D in the chelation step of protoporphyrin IX. Proc Natl Acad Sci USA 96:1941–1946

    Article  PubMed  CAS  Google Scholar 

  • Hukmani P, Tripathy BC (1992) Spectrofluorometric estimation of intermediates of chlorophyll biosynthesis: protoporphyrin IX, Mg-Protoporphyrin and Protochlorophyllide. Anal Biochem 206:125–130

    Article  PubMed  CAS  Google Scholar 

  • Hukmani P, Tripathy BC (1994) Chlorophyll biosynthetic reactions during senescence of excised barley (Hordeum vulgare L. cv IB 65) leaves. Plant Physiol 105:1295–1300

    PubMed  CAS  Google Scholar 

  • Jilani A, Kar S, Bose S, Tripathy BC (1996) Regulation of the carotenoid content and chloroplast development by levulinic acid. Physiol Plant 96:139–145

    Article  CAS  Google Scholar 

  • Johanningmeier U, Howell SH (1984) Regulation of Light-harvesting Chlorophyll-binding Protein mRNA Accumulation in Chlamydomonas reinhardi. J Biol Chem 259:13541–13549

    PubMed  CAS  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

    PubMed  CAS  Google Scholar 

  • Joyard J, Teyssier E, Miège C, Berny-Seigneurin D, Marèchal E, Block MA, Dorne AJ, Rolland N, Ajlani G, Douce R (1998) The biochemical machinery of plastid envelope membranes. Plant Physiol 118:715–723

    Article  PubMed  CAS  Google Scholar 

  • Kannangara CG, Andersen RV, Pontoppidan B, Willows R, von Wettstein D (1994) Enzymic and mechanistic studies on the conversion of glutamate to 5-aminolevulinate. In: Chadwick DJ, Ackrill K (eds) The biosynthesis of the tetrapyrrole pigments, Ciba Foundation Symposium 180. Wiley, England, pp 3–20

    Chapter  Google Scholar 

  • Kannangara CG, Vothknecht UC, Hansson M, von Wettstein D (1997) Magnesium chelatase: association with ribosomes and mutant complementation studies identify subunit Xantha-G as a functional counterpart of the Rhodobacter subunit BchD. Mol Gen Genet 254:85–92

    Article  PubMed  CAS  Google Scholar 

  • Keegstra K, Yousuf AE (1986) Isolation and characterization of chloroplast envelope membranes. Methods Enzymol 118:316–325

    CAS  Google Scholar 

  • King TE (1967) Preparation of succinate-cytochrome C reductase and the cytochrome b-c1,particle and reconstitution of succinate cytochrome C reductase. Methods Enzymol 10:216–225

    Article  CAS  Google Scholar 

  • Kropat J, Oster U, Rüdiger W, Beck CF (1997) Chlorophyll precursors are signals of chloroplast origin involved in light induction of nuclear heat-shock genes. Proc Natl Acad Sci USA 94:14168–14172

    Article  PubMed  CAS  Google Scholar 

  • Kropat J, Oster U, Rüdiger W, Beck CF (2000) Chloroplast signalling in the light induction of nuclear HSP70 genes requires the accumulation of chlorophyll precursors and their accessibility to cytoplasm/nucleus. Plant J 24:523–531

    Article  PubMed  CAS  Google Scholar 

  • Lee HJ, Ball MD, Parham R, Rebeiz CA (1992) Chloroplast biogenesis 65: Enzymic conversion of protoporphyrin IX to Mg-protoporphyrin IX in a subplastidic membrane fraction of cucumber etiochloroplasts. Plant Physiol 99:1134–1140

    PubMed  CAS  Google Scholar 

  • Lermontova I, Kruse E, Mock HP, Grimm B (1997) Cloning and characterization of a plastidal and a mitochondrial isoform of tobacco protoporphyrinogen IX oxidase. Proc Natl Acad Sci USA 94:8895–8900

    Article  PubMed  CAS  Google Scholar 

  • Li HM, Moore T, Keegstra K (1991) Targeting of proteins to the outer envelope membrane uses a different pathway than transport into chloroplasts. Plant Cell 3:709–717

    Article  PubMed  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  • Manohara MS, Tripathy BC (2000) Regulation of Protoporphyrin IX biosynthesis by intraplastidic compartmentalization and adenosine triphosphate. Planta 212:52–59

    Article  PubMed  CAS  Google Scholar 

  • Matile P, Hörtensteiner S, Thomas H, Kräutler B (1996) Chlorophyll breakdown in senescent leaves. Plant Physiol 112:1403–1409

    PubMed  CAS  Google Scholar 

  • Matile P, Hörtensteiner S, Thomas H (1999) Chlorophyll degradation. Annu Rev Plant Physiol Plant Mol Biol 50:67–95

    Article  PubMed  CAS  Google Scholar 

  • Matringe M, Camadro JM, Block MA, Joyard J, Scalla R, Labbe P, Douce R (1992) Localization within chloroplasts of protoporphyrinogen oxidase, the target enzyme for Diphenylether like herbicides. J Biol Chem 267:4646-4651

    PubMed  CAS  Google Scholar 

  • Mohapatra A, Tripathy BC (2002) Detection of protoporphyrin IX in envelope membranes of pea chloroplasts. Biochem Biophys Res Commun 299:751–754

    Article  PubMed  CAS  Google Scholar 

  • Mohapatra A, Tripathy BC (2003) Developmental changes in sub-plastidic distribution of chlorophyll biosynthetic intermediates in cucumber (Cucumis sativus L). J Plant Physiol 160:9–15

    Article  PubMed  CAS  Google Scholar 

  • Nakayama M, Masuda T, Sato N, Yamagata H, Bowler C, Ohta H, Shioi Y, Takamiya K (1995) Cloning, subcellular localization and expression of CHL I, a subunit of magnesium-chelatase in soybean. Biochem Biophys Res Commun 215:422–428

    Article  PubMed  CAS  Google Scholar 

  • Nakayama M, Masuda T, Bando T, Yamagata H, Ohta H, Takamiya K (1998) Cloning and expression of the soybean chl H gene encoding a subunit of Mg-chelatase and localization of the Mg2+ concentration-dependent Chl H protein within the chloroplast. Plant Cell Physiol 39:275–284

    PubMed  CAS  Google Scholar 

  • Papenbrock J, Gräfe S, Kruse E, Hänel F, Grimm B (1997) Mg-chelatase of tobacco: identification of a Chl D cDNA sequence encoding a third subunit, analysis of the interaction of the three subunits with the yeast two-hybrid system, and reconstitution of the enzyme activity by co-expression of recombinant CHL D, CHL H and CHL I. Plant J 12:981–990

    Article  PubMed  CAS  Google Scholar 

  • Pineau B, Dubertret G, Joyard J, Douce R (1986) Fluorescence properties of the envelope membranes from spinach chloroplasts: Detection of protochlorophyllide. J Biol Chem 261:9210–9215

    PubMed  CAS  Google Scholar 

  • Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975:384–394

    Article  CAS  Google Scholar 

  • Rebeiz CA, Matheis JR, Smith BB, Rebeiz CC, Dayton DF (1975) Chloroplast biogenesis: biosynthesis and accumulation of Mg-protoporphyrin IX monoester and longer wavelength metalloporphyrins by greening cotyledons. Arch Biochem Biophys 166:446–465

    Article  PubMed  CAS  Google Scholar 

  • Rebeiz CA, Montazer-Zouhoor A, Hopen HJ, Wu SM (1984) Photodynamic herbicides. I. Concept and phenomenology. Enzyme Microbiol Technol 6:390–401

    Article  CAS  Google Scholar 

  • Tewari AK, Tripathy BC (1998) Temperature-stress-induced impairment of chlorophyll biosynthetic reactions in cucumber (Cucumis sativus L) and wheat (Triticum aestivum L). Plant Physiol 117:851–858

    Article  CAS  Google Scholar 

  • Tewari AK, Tripathy BC (1999) Acclimation of chlorophyll biosynthetic reactions to temperature stress in cucumber (Cucumis sativus L). Planta 208:431–437

    Article  CAS  Google Scholar 

  • Tripathy BC, Rebeiz CA (1985) Chloroplast biogenesis: quantitative determination of monovinyl and divinyl Mg-protoporphyrins and protochlorophyllides by spectrofluorometry. Analytical Biochem 149:43–61

    Article  CAS  Google Scholar 

  • Tripathy BC, Rebeiz CA (1986) Chloroplast biogenesis: demonstration of monovinyl and divinyl monocarboxylic routes of chlorophyll biosynthesis in higher plants. J Biol Chem 261:13556–13564

    PubMed  CAS  Google Scholar 

  • Tripathy BC, Rebeiz CA (1988) Chloroplast Biogenesis 60: conversion of divinyl protochlorophyllide to monovinyl protochlorophyllide in greening barley, a dark monovinyl/ light divinyl plant species. Plant Physiol 87:89–94

    Article  PubMed  CAS  Google Scholar 

  • von Wettstein D, Gough S, Kannangara CG (1995) Chlorophyll biosynthesis. Plant Cell 7:1039–1057

    Article  Google Scholar 

  • Walker CJ, Willows RD (1997) Mechanism and regulation of Mg-chelatase. Biochem J 327:321–333

    PubMed  CAS  Google Scholar 

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Acknowledgments

We wish to thank Kenneth Keegstra, East Lansing, USA for OM 14 antibodies and Dr. A Matto, Beltsville, USA for LHCPII antibodies.

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Authors and Affiliations

  1. School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Anasuya Mohapatra & Baishnab C. Tripathy

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  1. Anasuya Mohapatra
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  2. Baishnab C. Tripathy
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Correspondence to Baishnab C. Tripathy.

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This work was supported by a grant from the Council of Scientific and Industrial Research (38/1079/03/EMRII) to BCT.

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Mohapatra, A., Tripathy, B.C. Differential distribution of chlorophyll biosynthetic intermediates in stroma, envelope and thylakoid membranes in Beta vulgaris . Photosynth Res 94, 401–410 (2007). https://doi.org/10.1007/s11120-007-9209-6

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  • DOI: https://doi.org/10.1007/s11120-007-9209-6

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