Abstract
The nuclear gene mutant chlorina-104 of barley (Hordeum vulgare L.) is chlorophyll-deficient when grown under high irradiance, particularly at low temperatures. Chlorina- 104 chloroplasts had fewer thylakoids than the wild type, and fewer appressed lamellae relative to non-appressed lamellae. The freeze-fracture ultrastructure showed a loss of particles from the protoplasmic fracture face of the stacked thylakoid region (PFs), consistent with the loss of most of the light-harvesting complex (LHC) II, and a loss of some of the large particles from the same face of the unstacked thylakoid region (PFu), indicating a loss of photosystem-I particles. The mutant is remarkable for the high density of particles on the exoplasmic fracture face of the unstacked thylakoid region (EFu), levels of which fell to normal after transfer to low light. The chlorophyll deficiency was shown to be primarily caused by the loss of LHCII and LHCI-680, with the consequent loss of much of the chlorophyll (Chl) b and the xanthophylls neoxanthin and lutein. The use of a monoclonal antibody which recognises the 23-kDa polypeptide of LHCI-680, confirmed that it was severely depleted in chloroplasts from chlorina-104 grown under restrictive conditions. The 77 K fluorescence emission spectrum was characterised by a pronounced shoulder at 720 nm, arising from the photosystem-I reaction centre (CPI). Since fluorescence from CPI is normally quenched by LHCI-730, this indicates that LHCI-680 mediates excitation energy transfer between LHCI-730 and the reaction centre. After moving seedlings to permissive conditions, LHCII and LHCI-680 began to accumulate in the chlorotic leaves and the fluorescence emission spectrum resembled that of wild-type leaves. Measurement of the steady-state mRNA levels with specific Cab probes, showed no difference between wild type and mutant, indicating that control of LHCII and LHCI-680 accumulation was at a post-transcriptional level.
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Abbreviations
- Cab :
-
gene symbol for chlorophyll a/b-proteins
- Chl:
-
chlorophyll
- CP:
-
chlorophyll protein
- EFs:
-
exoplasmic fracture face, stacked thylakoid region
- EFu:
-
exoplasmic fracture face, unstacked thylakoid region
- Fm/Fo :
-
ratio of fluorescence yield with saturating light conditions and ground level fluorescence
- LHC:
-
light-harvesting complex
- PAGE:
-
polyacrylamide gel electrophoresis
- PFs:
-
protoplasmic fracture face, stacked thylakoid region
- PFu:
-
protoplasmic fracture face, unstacked thylakoid region
- PSI-200:
-
“native” PSI complex
- SDS:
-
sodium dodecyl sulphate
References
Allen, K.D., Duysen, M.E., Staehelin, L.A. (1988) Biogenesis of thylakoid membranes is controlled by light intensity in the conditional chlorophyll b-deficient CD3 mutant of wheat. J. Cell Biol. 107, 907–919
Arnon, D.I. (1949) Copper enzymes in isolated chloroplasts. Polyphenol oxidase in Beta vulgaris. Plant Physiol. 24, 1–15
Bassi, R., Simpson, D. (1987) Chlorophyll-protein complexes of barley photosystem I. Eur. J. Biochem. 163, 221–230
Bassi, R., Machold, O., Simpson, D. (1985) Chlorophyll-proteins of two photosystem I preparations from maize. Carlsberg Res. Commun. 50, 145–162
Bassi, R., Høyer-Hansen, G., Barbato, R., Giacometti, G.M., Simpson, D.J. (1987) Chlorophyll-proteins of the photosystem II antenna system. J. Biol. Chem. 262, 13333–13341
Berthold, D.A., Babcock, G.T., Yocum, C.F. (1981) A highly resolved oxygen-evolving photosystem II preparation from spinach thylakoid membranes. FEBS Lett. 134, 231–234
Di Paolo, M.L., Dal Belin Peruffo, A., Bassi, R. (1990) Immunological studies on chlorophyll-a/b proteins and their distribution in thylakoid membrane domains. Planta 181, 275–286
Dunahay, T.G., Staehelin, L.A., Seibert, M., Ogilvie, P.D., Berg, S.P. (1984) Structural, biochemical and biophysical characterization of four oxygen evolving photosystem II preparations from spinach. Biochim. Biophys. Acta 764, 179–193
Dunahay, T.G., Staehelin, L.A. (1986) Isolation and characterization of a new minor chlorophyll a/b protein complex (CP24) from spinach. Plant Physiol. 80, 429–434
Feinberg, A.P., Vogelstein, B. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13
Fling, S.P., Gregerson, D.S. (1986) Peptide and protein molecular weight determination by electrophoresis using a high molarity Tris buffer system without urea. Anal. Biochem. 155, 83–88
Gamier, J., Maroc, J., Guyon, D. (1986) Low-temperature fluorescence emission spectra and chlorophyll-protein complexes in mutants of Chlamydomonas reinhardtii: evidence for a new chlorophyll-a-protein complex related to Photosystem I. Biochim. Biophys. Acta 851, 395–406
Glick, R.E., Melis, A. (1988) Minimum photosynthetic unit size in System I and System II of barley chloroplasts. Biochim. Biophys. Acta 934, 151–155
Greene, B.A., Allred, D.R., Morishige, D.T., Staehelin, L.A. (1988a) Hierarchical response of light harvesting chlorophyll-proteins in a light-sensitive chlorophyll b-deficient mutant of maize. Plant Physiol. 87, 357–364
Greene, B.A., Staehelin, L.A., Melis, A. (1988b) Compensatory alterations in the photochemical apparatus of a photoregulatory, chlorophyll b-deficient mutant of maize. Plant Physiol. 87, 365–370
Hoffman, N.E., Pichersky, E., Malik, V.S., Castresana, C., Ko, K., Darr, S.C., Cashmore, A.R. (1987) A cDNA clone encoding a photosystem I protein with homology to photosystem II chlorophyll a/b-binding polypeptides. Proc. Natl. Acad. Sci. USA 84, 8844–8848
Høyer-Hansen, G., Hønberg, L.S., Simpson, D.J. (1985) Monoclonal antibodies used for the characterization of the two putative iron-sulphur centre proteins associated with photosystem I. Carlsberg Res. Commun. 50, 23–35
Høyer-Hansen, G., Bassi, R., Hønberg, L.S., Simpson, D.J. (1988) Immunological characterization of chlorophyll a/b-binding proteins of barley thylakoids. Planta 173, 12–21
Ish-Shalom, D., Ohad, I.: (1983) Organization of chlorophyll-protein complexes of photosystem I in Chlamydomonas reinhardii. Biochim. Biophys. Acta 722, 498–507
Kloppstech, K., Schweiger, H.G. (1976) In vitro translation of polyA RNA from Acetabularia. Cytobiologie 13, 394–400
Knoetzel, J., Braumann, T., Grimme, L.H. (1988) Pigment-protein complexes of green algae: improved methodological steps for the quantification of pigments in pigment-protein complexes derived from the green algae Chlorella and Chlamydomonas. J. Photochem. Photobiol. B Biol. 1, 475–491
Knoetzel, J., Simpson, D. (1990) Characterisation of a temperature-sensitive mutant of barley. In: Current research in photosynthesis, vol. II, pp. 867–870, Baltscheffsky, M., ed. Kluwer Academic Publishers, Netherlands
Lam, E., Ortiz, W., Malkin, R. (1984) Chlorophyll a/b proteins of photosystem I. FEBS Lett. 168, 10–14
Maniatis, T., Fritsch, E.F., Sambrook, J. (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Mullet, J.E., Burke, J.J., Arntzen, C.J. (1980) Chlorophyll proteins of photosystem I. Plant Physiol. 65, 814–822
Oelmüller, R. (1989) Photooxidative destruction of chloroplasts and its effect on nuclear gene expression and extraplastidic enzyme levels. Photochem. Photobiol. 49, 229–239
Pichersky, E., Green, B.R. (1990) The extended family of chlorophyll a/b-binding proteins of PSI and PSII. Current research in photosynthesis, vol. III, pp. 553–556, Baltscheffsky, M., ed. Kluwer Academic Publishers, Netherlands
Pichersky, E., Bernatzky, R., Tanksley, S.D., Breidenbach, R.W., Kausch, A.P., Cashmore, A.R. (1985) Molecular characterization and genetic mapping of two clusters of genes encoding chlorophyll a/b-binding proteins in Lycopersicon esculentum (tomato). Gene 40, 247–258
Pichersky, E., Hoffman, N.E., Bernatzky, R., Piechulla, B., Tanksley, S.D., Cashmore, A.R. (1987a) Molecular characterization and genetic mapping of DNA sequences encoding the Type I chlorophyll a/b-binding polypeptide of photosystem I in Lycopersicon esculentum (tomato). Plant Mol. Biol. 9, 205–216
Pichersky, E., Hoffman, N.E., Malik, V.S., Bernatzky, R., Tanksley, S.D., Szabo, L., Cashmore, A.R. (1978b) The tomato Cab-4 and Cab-5 genes encode a second type of Cab polypeptides localized in photosystem II. Plant Mol. Biol. 9, 109–120
Pichersky, E., Tanksley, S.D., Piechulla, B., Stayton, M.M., Dunsmuir, P. (1988) Nucleotide sequence and chromosomal location of Cab-7, the tomato gene encoding the Type II chlorophyll a/b-binding polypeptide of photosystem I. Plant Mol. Biol. 11, 69–71
Pichersky, E., Brock, T.G., Nguyen, D., Hoffman, N.E., Piechulla, B., Tanksley, S.D., Green, B.R. (1989) A new member of the Cab gene family: structure, expression and chromosomal location of Cab-8, the tomato gene encoding the Type III chlorophyll a/b-binding polypeptide of photosystem I. Plant Mol. Biol. 12, 257–270
Sambrook, J., Fritsch, E.F., Maniatis, T. (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Schägger, H., von Jagow, G. (1987) Tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1–100 kDa. Anal. Biochem. 166, 368–379
Schwartz, E., Pichersky, E. (1990) Sequence of two tomato nuclear genes encoding chlorophyll a/b-binding proteins of CP24, a PSII antenna component. Plant Mol. Biol. 15, 157–160
Simpson, D. (1979) Freeze-fracture studies on barley plastid membranes III. Location of the light-harvesting chlorophyll-protein. Carlsberg Res. Commun. 44, 305–336
Simpson, D. (1983) Freeze-fracture studies on barley plastid membranes VI. Location of the chlorophyll a-protein 1. Eur. J. Cell Biol. 31, 305–314
Simpson, D. (1988) Low temperature absorption spectroscopy of barley mutants. Gaussian deconvolution and fourth derivative analysis. Carlsberg Res. Commun. 53, 343–356
Simpson, D., von Wettstein, D. (1980) Macromolecular physiology of plastids XIV. Viridis mutants in barley: genetic, fluoroscopic and ultrastructural characterisation. Carlsberg Res. Commun. 45, 283–314
Simpson, D.J., Machold, O., Høyer-Hansen, G., von Wettstein, D. (1985) Chlorina mutants of barley (Hordeum vulgare L.). Carlsberg Res. Commun. 50, 223–238
Somerville, C.R. (1986) Analysis of photosynthesis with mutants of higher plants and algae. Annu. Rev. Plant Physiol. 37, 467–507
Staehelin, L.A. (1986) Chloroplast structure and supramolecular organization of photosynthetic membranes. In: Encyclopedia of Plant Physiology, vol. 19, pp. 1–84 Staehelin, L.A., Arntzen, C.J. eds. Springer-Verlag, Berlin
Taylor, W.C. (1989) Regulatory interactions between nuclear and plastid genomes. Annu Rev. Plant Physiol. 40, 211–233
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We wish to thank Professor Diter von Wettstein (Carlsberg Laboratory, Copenhagen Valby, Denmark) for his encouragement and advice during the course of these investigations, and for critically reading the manuscript. We are grateful for Cab clones sent to us by Dr. Eran Pichersky (University of Michigan, Ann Arbor, Mich., USA), who also communicated sequence data prior to publication. We thank Klaus N. Kristiansen (Carlsberg Laboratory) for his patient and expert guidance with the in-vitro transcriptions and Northern blots. We also thank Nina Rasmussen for drawing the figures, Ann-Sofi Steinholz for photography, and Jean Sage for thin-sectioning.
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Knoetzel, J., Simpson, D. Expression and organisation of antenna proteins in the light-and temperature-sensitive barley mutant chlorina-104 . Planta 185, 111–123 (1991). https://doi.org/10.1007/BF00194522
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DOI: https://doi.org/10.1007/BF00194522