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Screening of chlorina mutants of barley (Hordeum vulgare L.) with antibodies against light-harvesting proteins of PS I and PS II: Absence of specific antenna proteins

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Abstract

Twenty-three chlorina (clo) mutants from the barley mutant collection of the Carlsberg Laboratory, Copenhagen, were tested for the presence of the four light-harvesting chlorophyll (Chl) a/b-binding proteins (LHC) of Photosystem I (Lhca1-4) and the PS II antenna proteins Lhcb1-3 (LHC II), Lhcb4-6 (CP29, CP26, CP24) and PsbS (CP22) using monospecific and monoclonal antibodies. Mutants allelic to barley mutant clo-f2, impaired in Chl b synthesis, provided evidence that Lhca4, Lhcb1 and Lhcb6 are unstable in the absence of Chl b, and the accumulation of Lhcb2, Lhcb3 and Lhcb4 is also impaired. Mutants at the locus chlorina-a (clo-a117, clo-a126 and clo-a134) lack or have only trace amounts of Lhca1, Lhca4, Lhcb1 and Lhcb3, whereas a mutant at the locus chlorina-b (clo-b125) had reduced amounts of all Lhca proteins. These two mutations could have an effect in protein import or assembly. Evidence is presented that Lhcb5 is the innermost LHC protein of PS II, and that Lhca1 and Lhca4, which have been supposed to be intimately associated in the LHCI-730 complex, can accumulate independently of each other. 77 K fluorescence emission spectra taken from leaves of clo-f2 101, clo-a126 and clo-b125 indicate that chlorophyll(s) emitting at 742 nm are coupled to the presence of Lhca4 that is bound to the reaction centre, and those emitting around 730 nm are located on Lhca1.

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References

  • Adamska I (1995) Light stress proteins (ELIPs); the intriguing relatives of the cab gene family. In: Mathis P (ed) Photosynthesis: from Light to Biosphere, Vol III, pp 887-892. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Bassi R and Simpson DJ (1987) Chlorophyll-protein complexes of barley Photosystem I. Eur J Biochem 163: 221-230

    Google Scholar 

  • Bassi R, Soen SY, Frank G, Zuber H and Rochaix JD (1992) Characterization of chlorophyll a/b proteins of Photosystem I from Chlamydomonas reinhardtii. J Biol Chem 267: 25714-25721

    Google Scholar 

  • Bellemare G, Bartlett SG and Chua NH (1982) Biosynthesis of chlorophyll a/b-binding polypeptides in wild type and the chlorina-f2 mutant of barley. J Biol Chem 257: 7762-7767

    Google Scholar 

  • Boekema EJ, Wynn RM and Malkin R (1990) The structure of spinach Photosystem I studied by electron microscopy. Biochim Biophys Acta 1017: 49-56

    Google Scholar 

  • Bruce BD and Malkin R (1988) Subunit stoichiometry of the chloroplast Photosystem I complex. J Biol Chem 263: 7302-7308

    Google Scholar 

  • Burke JJ, Steinback KE and Arntzen CJ (1979) Analysis of the light-harvesting pigment-protein complex of wild type and a chlorophyll-b-less mutant of barley. Plant Physiol 63: 237-243

    Google Scholar 

  • Croce R, Zucchelli G, Garlaschi FM, Bassi R and Jennings RC (1996) Excited state equilibrium in Photosystem I-light-harvesting I complex: P700 is almost isoenergetic with its antenna. Biochemistry 35: 8572-8579

    Google Scholar 

  • Falbel TG and Staehelin LA (1992) Species-related differences in the electrophoretic behaviour of CP29 and CP26: An immunochemical analysis. Photosynth Res 34: 249-262

    Google Scholar 

  • Falbel TG and Staehelin LA (1994) Characterization of a family of chlorophyll-deficient wheat (Triticum) and barley (Hordeum vulgare) mutants with defects in the magnesium-insertion step of chlorophyll biosynthesis. Plant Physiol 104: 639-648

    Google Scholar 

  • Falbel TG, Staehelin LA and Adams III WW (1994) Analysis of xanthophyll cycle carotenoids and chlorophyll fluorescence in light intensity-dependent chlorophyll-deficient mutants of wheat and barley. Photosynth Res 42: 191-202

    Google Scholar 

  • Fling SP and Gregerson DS (1986) Peptide and protein molecular weight determination by electrophoresis using a high-molarity tris system without urea. Anal Biochem 155: 83-88

    Google Scholar 

  • Funk C, Schröder WP, Napiwotzki A, Tjus SE, Renger G and Andersson B (1995) The PS II-S protein of higher plants: A new type of pigment-binding protein. Biochemistry 34: 11133-11141

    Google Scholar 

  • Gobets B, van Amerongen H, Monshouwer R, Kruip J, Rögner M, van Grondelle R and Dekker JP (1994) Polarized site-selected fluorescence spectroscopy of isolated Photosystem I particles. Biochim Biophys Acta 1188: 75-85

    Google Scholar 

  • Hankamer B, Nield J, Zheleva D, Boekema E, Jansson S and Barber J (1997) Isolation and biochemical characterization of monomeric and dimeric PS II complexes from spinach and their relevance to the organization of Photosystem II in vivo. Eur J Biochem 243: 422-429

    Google Scholar 

  • Harrison MA, Nemson JA and Melis A (1993) Assembly of the chlorophyll a/b light-harvesting complex of barley (Hordeum vulgare L.): Immunochemical analysis of chlorophyll b-less and chlorophyll b-deficient mutants. Photosynth Res 38: 141-151

    Google Scholar 

  • Høyer-Hansen G, Bassi R, Hønberg LS and Simpson DJ (1988) Immunological characterization of chlorophyll a/b-binding proteins of barley thylakoids. Planta 173: 13-21

    Google Scholar 

  • Ikeuchi M, Hirano A and Inoue Y (1991) Correspondence of apoproteins of light-harvesting chlorophyll a/b complexes associated with Photosystem I to cab genes: Evidence for a novel type IV apoprotein. Plant Cell Physiol 32(1): 103-112

    Google Scholar 

  • Jansson S and Gustafsson P (1990) Type I and Type II genes for the chlorophyll a/b-binding protein in the gymnosperm Pinus sylvestris (Scots pine): cDNA cloning and sequence analysis. Plant Mol Biol 14: 287-296

    Google Scholar 

  • Jansson S and Gustafsson P (1991) Evolutionary conservation of the chlorophyll a/b-binding proteins: cDNAs encoding Type I, II and II LHC I polypeptides from the gymnosperm Scots pine. Mol Gen Genet 229: 67-76

    Google Scholar 

  • Jansson S, Pichersky E, Bassi R, Green BR, Ikeuchi M, Melis A, Simpson DJ, Spangfort M, Staehelin LA and Thornber JP (1992) A nomenclature for the genes encoding the chlorophyll a/b-binding proteins of higher plants. Plant Mol Biol Rep 10: 242-253

    Google Scholar 

  • Jansson S (1994) The light-harvesting chlorophyll a/b-binding proteins. Biochim Biophys Acta 1184: 1-19

    Google Scholar 

  • Jansson S, Andersen B and Scheller HV (1996) Nearest neighbor analysis of higher plant Photosystem I holocomplex. Plant Physiol 112: 409-420

    Google Scholar 

  • Knoetzel J and Simpson D (1991) Expression and organisation of antenna proteins in the light-and temperature-sensitive barley mutant chlorina-104. Planta 185: 111-123

    Google Scholar 

  • Knoetzel J, Svendsen I and Simpson DJ (1992) Identification of the Photosystem I antenna polypeptides in barley. Eur J Biochem 206: 209-215

    Google Scholar 

  • Krause GH and Weis E (1991) Chlorophyll fluorescence and photosynthesis: The basics. Ann Rev Plant Physiol Plant Mol Biol 42: 313-349

    Google Scholar 

  • Król M, Spangfort MD, Huner NPA, Öquist G, Gustafsson P and Jansson S (1995) Chlorophyll a/b-binding proteins, pigment conversions, and early light-induced proteins in a chlorophyll b-less barley mutant. Plant Physiol 107: 873-883

    Google Scholar 

  • Ljungberg U, Åkerlund HE and Andersson B (1986) Isolation and characterization of the 10-kDa and 22-kDa polypeptides of higher plant Photosystem II. Eur J Biochem 158: 477-482

    Google Scholar 

  • Machold O, Meister A, Sagromsky H, Høyer-Hansen G and von Wettstein D (1977) Composition of photosynthetic membranes of wild-type barley and chlorophyll b-less mutants. Photosynthetica 11(2): 200-206

    Google Scholar 

  • Machold O (1981) Thylakoid membrane proteins of higher plant chloroplasts. Biochem Physiol Pflanzen 176: 535-548

    Google Scholar 

  • Mullet JE, Burke JJ and Arntzen CJ (1980) Chlorophyll proteins of Photosystem I. Plant Physiol 65: 814-822

    Google Scholar 

  • Nielsen VS, Mant A, Knoetzel J, Møller BL and Robinson C (1994) Import of barley Photosystem I subunit N into the thylakoid lumen is mediated by a bipartite presequence lacking an intermediate processing site. Role of ΔpH in translocation across the thylakoid membrane. J Biol Chem 269: 3762-3766

    Google Scholar 

  • Nielsen VS, Scheller HV and Møller BL (1996) The Photosystem I mutant viridis-zb 63 of barley (Hordeum vulgare) contains low amounts of active but unstable Photosystem I. Physiol Plant 98: 637-644

    Google Scholar 

  • Paulsen H (1995) Chlorophyll a/b-binding proteins. Photochem Photobiol 62: 367-382

    Google Scholar 

  • Peter GF and Thornber JP (1991a) Biochemical evidence that the plant Photosystem II core complex is organized as a dimer. Plant Cell Physiol 32: 1237-1250

    Google Scholar 

  • Peter GF and Thornber JP (1991b) Biochemical composition and organization of higher plant Photosystem II light-harvesting pigment proteins. J. Biol. Chem 266: 16745-16754

    Google Scholar 

  • Preiss S and Thornber JP (1995) Stability of the apoproteins of light-harvesting complex I and II during biogenesis of thylakoids in the chlorophyll b-less barley mutant Chlorina f2. Plant Physiol 107: 709-717.

    Google Scholar 

  • Ryrie IJ (1983) Immunological evidence for apoproteins of the light-harvesting chlorophyll-protein complex in a mutant of barley lacking chlorophyll b. Eur J Biochem 131: 149-155

    Google Scholar 

  • Shubin VV, Bezsmertnaya IN and Karapetyan NV (1992) Isolation from Spirulina membranes of two Photosystem I-type complexes, one of which contains chlorophyll responsible for the 77 K fluorescence band at 760 nm. FEBS Lett 309: 340-342

    Google Scholar 

  • Shubin VV, Tsuprun VL, Bezsmertnaya IN and Karapetyan NV (1993) Trimeric forms of Photosystem I reaction center complexes pre-exist in the membranes of the cyanobacterium Spirulina platensis. FEBS Lett 334: 79-82

    Google Scholar 

  • Sigrist M and Staehelin LA (1992) Identification of type 1 and type 2 light-harvesting chlorophyll a/b-binding proteins using monospecific antibodies. Biochim Biophys Acta 1098: 191-200

    Google Scholar 

  • Simpson DJ (1979) Freeze-fracture studies on barley plastid membranes III. Localisation of the light-harvesting chlorophyll protein. Carlsberg Res Commun 44: 305-336

    Google Scholar 

  • Simpson DJ, Machold O, Høyer-Hansen G and von Wettstein D (1985) Chlorina mutants of barley (Hordeum vulgare L.). Carlsberg Res Commun 50: 223-238

    Google Scholar 

  • Simpson DJ and v Wettstein D (1992) Nuclear genes affecting the chloroplast. Stock list of mutants kept at the Carlsberg Laboratory. Barley Genetics Newsletter 21: 102-108

    Google Scholar 

  • Simpson DJ and Knoetzel J (1996) Light-harvesting complexes in plants and algae: Introduction, survey and nomenclature. In: Ort RO and Yocum CF (eds) Oxygenic Photosynthesis: The Light Reactions, pp 493-506. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Thornber JP and Highkin HR (1974) Composition of the photosynthetic apparatus of normal barley leaves and a mutant lacking chlorophyll b. Eur J Biochem 41: 109-116

    Google Scholar 

  • Tjus SE, Roobol-Bóza M, Pålsson LO and Andersson B (1995) Rapid isolation of Photosystem I chlorophyll-binding proteins by anion exchange perfusion chromatography. Photosynth Res 45: 41-49

    Google Scholar 

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

    Google Scholar 

  • White MJ and Green BR (1987) Polypeptides belonging to each of the three major chlorophyll a+b protein complexes are present in a chlorophyll b-less barley mutant. Eur J Biochem 165: 531-535

    Google Scholar 

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Authors and Affiliations

  1. Institute of Cell Biology, Biochemistry and Biotechnology, University of Bremen, Leobener Straße/NW II, 28334, Bremen, Germany

    Björn Bossmann & Jürgen Knoetzel

  2. Department of Plant Physiology, Ume University, 90187, Ume, Sweden

    Stefan Jansson

Authors
  1. Björn Bossmann
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  2. Jürgen Knoetzel
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  3. Stefan Jansson
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Bossmann, B., Knoetzel, J. & Jansson, S. Screening of chlorina mutants of barley (Hordeum vulgare L.) with antibodies against light-harvesting proteins of PS I and PS II: Absence of specific antenna proteins. Photosynthesis Research 52, 127–136 (1997). https://doi.org/10.1023/A:1005823711838

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