Abstract
Thirty-four recessive photosynthetic mutants of the high-chlorophyll-fluorescence (hcf) phenotype have been isolated by screening 7700 M2 progenies of ethyl methane sulfonate-treated seeds ofArabidopsis thaliana. Most of the mutants isolated were found to be seedlinglethal, but could be grown on sucrose-supplemented media. Chlorophyll (Chl) fluorescence induction, absorption changes in the reaction-centre chlorophyll of PS I (P700) at 830 nm and Chla/Chlb ratios were recorded in order to probe the photosynthetic functions and to define the mutational lesion. These studies were complemented by immunoblot and Northern analyses which finally led to the classification of the mutants into six different groups. Four classes of mutants were affected in PS I, PS II (two different classes) or the intersystem electron-transport chain, respectively. A fifth mutant class was of pleiotropic nature and the sixth class comprised a Chlb-deficient mutant. Several of the mutants showed severe deficiencies in the levels of subunits of PS I, PS II or the cytochromeb 6/f complex. Thus the mutational lesions could be located precisely. Only one mutant was defective in the transcript patterns of some plastid-encoded photosynthesis genes. Hence most of the mutants isolated appear to be affected in translational and post-translational regulatory processes of thylakoid membrane biogenesis or in structural genes encoding constituent subunits of the thylakoid protein complexes.
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Abbreviations
- Chl:
-
chlorophyll
- ΔA:
-
in-vivo absorbance change of P700 at 830 nm measured immediately after interruption of the actinic white light which had been given for four minutes
- ΔAmax:
-
in-vivo absorbance change of P700 at 830 nm induced by background far-red light at 720 nm
- EMS:
-
ethyl methane sulfonate
- Fo and Fm :
-
minimal and maximal Chla fluorescence of dark-adapted leaves
- F′o, F′m and Fs :
-
minimal, maximal and steady-state Chla fluorescence of light-adapted leaves
- Fv :
-
variable fluorescence
- ΦI:
-
quantum yield of PS I
- ΦII:
-
quantum yield of PS II
- hcf :
-
high chlorophyll fluorescence
- P700:
-
reaction-centre chlorophyll of PS I
- qN :
-
non-photochemical quenching
- qp :
-
photochemical quenching
References
Aristarkkhov AI, Nikandrov VV, Krasnovskii AA (1987) Ascorbate permeability of chloroplast thylakoid membrane; reduction of plastoquinones. Biokhimiya 52: 2051–2060
Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase inBeta vulgaris. Plant Physiol 24: 1–13
Arnon DI, Barber J (1990) Photoreduction of NADP+by isolated reaction centers of photosystem II: Requirement for plastocyanin. Proc Natl Acad Sci USA 87: 5930–5934
Asada K, Heber U, Schreiber U (1993) Electron flow to the intersystem chain from stromal components and cyclic electron flow in maize chloroplasts, as detected in leaves by monitoring redox change of P700 and chlorophyll fluorescence. Plant Cell Physiol 34: 39–50
Barkan A (1993) Nuclear mutants of maize with defects in chloroplast polysome assembly have altered chloroplast RNA metabolism. Plant Cell 5: 389–402
Barkan A, Miles D, Taylor WC (1986) Chloroplast gene expression in nuclear, photosynthetic mutants of maize. EMBO J 5: 1421–1427
Barkan A, Voelker R, Mendel-Hartvig J, Johnson D, Walker M (1995) Genetic analysis of chloroplast biogenesis in higher plants. Physiol Plant 93: 163–170
Bennoun P (1982) Evidence for a respiratory chain in the chloroplast. Proc Natl Acad Sci USA 79: 4352–4356
Bennoun P, Delepelaire P (1982) Isolation of photosynthesis mutants inChlamydomonas. In: Edelman M, Hallick RB, Chua NH (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, Amsterdam, pp 25–38
Choquet Y, Goldschmidt-Clermont M, Girard-Bascou J, Kück U, Bennoun P, Rochaix JD (1988) Mutant phenotypes support a transsplicing mechanism for the expression of the tripartitepsaA gene in theC. reinhardtii chloropla0st. Cell 52: 903–913
Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995
Demmig-Adams B (1990) Carotenoids and photoprotection in plants: A role for the xanthophyll zeaxanthin. Biochim Biophys Acta BioEnerget 1020: 1–24
de Vitry C, Olive J, Drapier D, Recouvreur M, Wollman F-A (1989) Posttranslational events leading to the assembly of photosystem II protein complex: A study using photosynthesis mutants fromChlamydomonas reinhardtii. J Cell Biol 109: 991–1006
Dinkins RD, Bandaranayake H, Green BR, Griffiths AJF (1994) A nuclear photosynthetic electron transport mutant ofArabidopsis thaliana with altered expression of the chloroplastpetA gene. CurrGenet 25: 282–288
Drapier D, Girard-Bascou J, Wollman F-A (1992) Evidence for nuclear control of the expression of theatpA andatpB chloroplast genes inChlamydomonas. Plant Cell 4: 283–295
Erickson JM, Rochaix JD (1992) The molecular biology of photosystemII. In: Barber J (ed) Topics in photosynthesis, Vol 11: The photosystems: Structure, function and molecular biology. Elsevier Science Publishers, Amsterdam, pp 101–177
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13
Feldmann KA (1991) T-DNA insertion mutagenesis inArabidopsis: mutational spectrum. Plant J 1: 71–82
Gamier J, Guyon D, Picaud D (1979) Characterization of new strains of nonphotosynthetic mutants ofChlamydomonas reinhardtii. I. Fluorescence, photochemical activities, chlorophyll-protein complexes. Plant Cell Physiol 20: 1013–1027
Genty B, Briantais J-M, Baker NR (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990: 87–92
Groom QJ, Kramer DM, Crofts AR, Ort D (1993) The nonphotochemical reduction of plastoquinone in leaves. Photosynth Res 36: 205–215
Harbinson J, Woodward FI (1987) The use of light-induced absorbance changes at 820 nm to monitor the oxidation state of P700 in leaves. Plant Cell Environ 10: 131–140
Harris GC, Heber U (1993) Effects of anaerobiose on chlorophyll fluorescence yield in spinach (Spinacia oleracea) leaf discs. Plant Physiol 101: 1169–1173
Hiller RG, Møller BL, Høyer-Hansen G (1980) Characterization of six putative photosystem I mutants in barley. Carlsberg Res Commun 45: 315–328
Höfer MU, Santore UJ, Westhoff P (1992) Differential accumulation of the 10-, 16- and 23-kDa peripheral components of the water-splitting complex of photosystem II in mesophyll and bundle-sheath chloroplasts of the dicotyledonous C4 plantFlaveria trinerma (Spreng.) C. Mohr. Planta 186: 304–312
Horton P, Ruban AV (1992) Regulation of photosystem II. Photosynth Res 34: 375–385
Jürgens G, Mayer U, Torres Ruiz RA, Berleth T, Misra S (1991) Genetic analysis of pattern formation in theArabidopsis embryo. Development Suppl 1: 27–38
Juhler RK, Miller M, Simpson D, Cox RP (1993) Chlorophyll fluorescence transients in a barley mutant lacking photosystem I. Photosynth Res 35: 305–310
Kuchka MR, Goldschmidt-Clermont M, van Dillewijn J, Rochaix J-D (1989) Mutation at theChlamydomonas nuclear NAC2 locus specifically affects stability of the chloroplastpsbD transcript encoding polypeptide D2 of PS II. Cell 58: 869–876
Kyhse-Andersen J (1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide gels to nitrocellulose. J Biochem Biophys Methods 10: 203–209
Lotan O, Streubel M, Westhoff P, Nechushtai R (1993) Subunit III (Psa-F) of photosystem I reaction center of the C4 dicotyledonFlaveria trinerma. Plant Mol Biol 21: 573–577
Mäenpää P, Aro E-M, Somersalo S, Tyystjärvi E (1988) Rearrangement of the chloroplast thylakoid at chilling temperature in the light. Plant Physiol 87: 762–766
Meierhoff K, Westhoff P (1993) Differential biogenesis of photosystem II in mesophyll and bundle-sheath cells of mono cotyledonous NADP-malic enzyme-type C4 plants: The non-stoichiometric abundance of the subunits of photosystem II in the bundle-sheath chloroplasts and the translational activity of the plastome-encoded genes. Planta 191: 23–33
Melis A (1991) Dynamics of photosynthetic membrane composition and function. Biochim Biophys Acta Bio-Energet 1058: 87–106
Mi H, Endo T, Schreiber U, Ogawa T, Asada K (1992) Electron donation from cyclic and respiratory flows to the photosynthetic intersystem chain is mediated by pyridine nucleotide dehydrogenase in the cyanobacteriumSynechocystis PCC 6803. Plant Cell Physiol 33: 1233–1237
Miles D (1980) Mutants of higher plants: maize. Methods Enzymol 69: 3–23
Miles D (1982) The use of mutations to probe photosynthesis in higher plants. In: Edelman M, Hallick RB, Chua NH (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, New York, pp 75–107
Mohr H (1986) Control by light of plastidogenesis as part of a control system. In: Akoyunoglou G, Senger H (eds) Regulation of chloroplast differentiation. Alan R Liss, pp 623–634
Mullet JE (1988) Chloroplast development and gene expression. Annu Rev Plant Physiol Plant Mol Biol 39: 475–502
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15: 473–497
Nikaido H, Saier MH Jr (1992) Transport proteins in bacteria: Common themes in their design. Science 258: 936–942
Ohyama K (1992) Organization and expression of genes of plastid chromosomes from non-angiospermous land plants and green algae.In: Herrmann RG (ed.) Plant gene research: Cell organelles. Springer Verlag, Wien-New York, pp 137–163
Oswald A, Streubel M, Ljungberg U, Hermans J, Eskins K, Westhoff P (1990) Differential biogenesis of photosystem II in mesophyll and bundle sheath cells of NADP malic enzyme-type C4 plants. A comparative protein and RNA analysis. Eur J Biochem 190: 185–194
Peltier G, Schmidt GW (1991) Chlororespiration: An adaptation to nitrogen deficiency inChlamydomonas reinhardtii. Proc Natl Acad Sci USA 88: 4791–4795
Pepper A, Delaney TP, Chory J (1993) Genetic interactions in plant photomorphogenesis. Sem Dev Biol 4: 15–22
Rédei GP (1965) Non mendelian megagametogenesis inArabidopsis. Genetics 51: 857–872
Rédei GP, Koncz C (1992) Classical mutagenesis. In: Koncz C, Chua NH, Schell J (eds) Methods in Arabidopsis research. World Scientific, Singapore, pp 16–82
Rees D, Horton P (1990) The mechanisms of changes in photosystem II efficiency in spinach thylakoids. Biochim Biophys Acta 1016: 219–227
Schägger H, von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166: 368–379
Schreiber U (1986) Detection of rapid induction kinetics with a new type of high-frequency modulated chlorophyll fluorometer. Photosynth Res 9: 261–272
Schreiber U, Klughammer C, Neubauer C (1988) Measuring P700 absorbance changes around 800 nm with a new type of pulse modulation system. Z Naturforsch 43C: 686–698
Sieburth LE, Berry-Lowe S, Schmidt GW (1991) Chloroplast RNA stability inChlamydomonas: Rapid degradation ofpsbB andpsbC transcripts in two nuclear mutants. Plant Cell 3: 175–189
Siefermann-Harms D (1987) The light harvesting and protective function of carotenoids in photosynthetic membranes. Physiol Plant 69: 561–568
Simpson DJ, Robinson SP (1984) Freeze fracture ultrastructure of thylakoid membranes in chloroplasts from manganese-deficient plants. Plant Physiol 74: 735–741
Simpson DJ, 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 DJ, Machold O, Hqyer-Hansen G, von Wettstein D (1985) Chlorina mutants of barley (Hordeum vulgare L.). Carlsberg Res Commun 50: 223–238
Somerville CR (1986) Analysis of photosynthesis with mutants of higher plants and algae. Annu Rev Plant Physiol 37: 467–507
Somerville CR, Ogren WL (1982) Isolation of photorespiration mutants inArabidopsis thaliana. In: Edelman M, Hallick RB, Chua NH (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, New York, pp 129–139
Susek RE, Ausubel FM, Chory J (1993) Signal transduction mutants of arabidopsis uncouple nuclear CAB and RBCS gene expression from chloroplast development. Cell 74: 787–799
Taylor WC (1989) Regulatory interactions between nuclear and plastid genomes. Annu Rev Plant Physiol Plant Mol Biol 40: 211–233
Taylor WC, Barkan A, Martienssen RA (1987) Use of nuclear mutants in the analysis of chloroplast development. Dev Genet 8: 305–320
Thompson KL, Brudvig GW (1988) Cytochromeb-559 may function to protect photosystem II from photoinhibition. Biochemistry 27: 6653–6658
van Kooten O, Snel JFH (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth Res 25: 147–150
Vermaas W (1993) Molecular-biological approaches to analyze photosystem II structure and function. Annu Rev Plant Physiol Plant Mol Biol 44: 457–481
Walbot V (1992) Strategies for mutagenesis and gene cloning using transposon tagging and T-DNA insertional mutagenesis. Annu Rev Plant Physiol Plant Mol Biol 43: 49–82
Westhoff P, Herrmann RG (1988) Complex RNA maturation in chloroplasts. ThepsbB operon from spinach. Eur J Biochem 171: 551–564
Westhoff P, Jansson C, Klein-Hitpaß L, Berzborn R, Larsson C, Bartlett SG (1985) Intracellular coding sites of polypeptides associated with photosynthetic oxygen evolution of photosystem II. Plant Mol Biol 4: 137–146
Westhoff P, Offermann-Steinhard K, Höfer M, Eskins K, Oswald A, Streubel M (1991) Differential accumulation of plastid transcripts encoding photosystem II components in the mesophyll and bundlesheath cells of monocotyledonous NADP-malic enzyme-type C4 plants. Planta 184: 377–388
Whittier RF, Sugiura M (1992) Plastid chromosomes from vascular plants — genes. In: Herrmann RG (ed.) Plant gene research: Cell organelles. Springer Verlag, Wien-New York, pp 164–182
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This work was supported by grants from the Deutsche Forschungs-gemeinschaft (SFB 189) and the Fonds der Chemischen Industrie. We are very grateful to W. Rogmann and coworkers for the careful cultivation of the plants in the greenhouse. We thank Dr. P. Jahns (Institut für Biochemie der Pflanzen, Universität Düsseldorf, Germany) for the critical reading of the manuscript. Parts of this work have been submitted for the doctoral theses of Dr. K. Meierhoff and J. Meurer.
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Meurer, J., Meierhoff, K. & Westhoff, P. Isolation of high-chlorophyll-fluorescence mutants ofArabidopsis thaliana and their characterisation by spectroscopy, immunoblotting and Northern hybridisation. Planta 198, 385–396 (1996). https://doi.org/10.1007/BF00620055
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DOI: https://doi.org/10.1007/BF00620055