Abstract
A shift in the ratio of chlorophyll (Chl) a and Chl b is an early indicator of heat bleaching in Euglena gracilis. This observation prompted us to consider whether or not changes in steady-state levels of chloroplast transcripts and in transcriptional activity could limit the synthesis of Chl a-binding proteins in bleaching plastids. We found that the mature transcripts for CP47 and CP43, the Chl a-binding apoproteins of the proximal antenna of photosystem II, decline sharply very early during bleaching. Our study also shows that transcription of psbB and psbC, the chloroplast genes encoding CP47 and CP43, remains essentially unchanged during the same interval. We conclude that posttranscriptional events, such as mRNA stability, could play a major role in initiating an irreversible loss of chloroplast function in Euglena at a moderately elevated temperature. Lack of these transcripts would eventually impair the assembly of photosystem II in thylakoids.
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References
Al-khatib K, Paulsen GM: Enhancement of thermal injury to photosynthesis in wheat plants and thylakoids by high light intensity. Plant Physiol 90: 1041–1048 (1989).
Armond PA, Björkman O, Staehelin LA: Dissociation of supramolecular complexes in chloroplast membranes. A manifestation of heat damage to the photosynthetic apparatus. Biochim Biophys Acta 601: 433–442 (1980).
Armond PA, Schreiber U, Björkman O: Photosynthetic acclimation to temperature in the desert shrub, Larrea divaricata. Light-harvesting efficiency and electron transport. Plant Physiol 61: 411–415 (1978).
Arnon DI: Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24: 1–15 (1949).
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K: Current Protocols in Molecular Biology. John Wiley, New York (1987).
Berry J, Björkman O: Photosynthetic response and adaptation to temperature in higher plants. Annu Rev Plant Physiol 31: 491–543 (1980).
Brandt P, Wiessner W: Different temperature optima of the DNA-dependent RNA polymerases of Euglena gracilis, strain Z and their role for the experimental bleaching by higher temperature. Z Pflanzenphysiol 85: 53–60 (1977).
Bricker TM: The structure and function of CPa-1 and CPa-2 in photosystem II. Photosynth Res 24: 1–13 (1990).
Chomczynski P, Sachi N: Single-step method of RNA isolation by thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159 (1987).
Conkling BA, Thomas EJ, Ortiz W: Delayed but complete loss of chloroplast DNA in heat-bleaching cultures of Euglena gracilis. J Plant Physiol 142: 307–311 (1993).
Cushman JC, Hallick RB, Price CA: The two genes for the P700 chlorophyll a apoproteins on the Euglena gracilis chloroplast genome contain multiple introns. Curr Genet 13: 159–171 (1988).
Eichacker L, Paulsen H, Rüdiger W: Synthesis of chlorophyll a regulates translation of chlorophyll a apoproteins P700, CP47, CP43 and D2 in barley etioplasts. Eur J Biochem 205: 17–24 (1992).
Eichacker LA, Soll J, Lauterbach P, Rüdiger W, Klein RR, Mullet JE: In vitro synthesis of chlorophyll a in the dark triggers accumulation of chlorophyll a apoproteins in barley etioplasts. J Biol Chem 265: 13566–13571 (1990).
Falk J, Schmidt A, Krupinska K: Characterization of plastid DNA transcription in ribosome deficient plastids of heat-bleached barley leaves. J Plant Physiol 141: 176–181 (1993).
Feierabend J: Inhibition of chloroplast ribosome formation by heat in higher plants. In: Edelman M, Hallick RB, Chua NH (eds) Methods in Chloroplast Molecular Biology, pp, 671–680. Elsevier, Amsterdam (1982).
Flamant F, Heizmann P, Nigon V: Rearrangement of chloroplast ribosomal cistrons by unequal crossing-over in Euglena gracilis. Curr Genet 8: 9–13 (1984).
Hallick RB, Gingrich JC, Johanningmeier U, Passavant CW: Introns in Euglena and Nicotiana chloroplast protein genes. In: VanVloten-Doting L, Groot GSP, Hall TC (eds) Molecular Form and Function of the Plant Genome, pp. 211–220. Plenum, New York (1985).
Hallick RB, Spielmann A, Stutz E: Complete sequence of Euglena gracilis chloroplast DNA. Nucl Acids Res 21: 3537–3544 (1993).
Herrmann RG, Feierabend J: The present of DNA in ribosome-deficient plastids of heat-bleached bye leaves. Eur J Biochem 104: 603–609 (1980).
Hess WR, Prombona A, Fieder B, Subramanian AR, Börner T: Chloroplast rps15 and the rpoB/C1/C2 gene cluster are strongly transcribed in ribosome-deficient plastids: evidence for a functioning non-chloroplastencoded RNA polymerase EMBO J 12: 563–571 (1993).
Hollingsworth MJ, Johanningmeier U, Karabin GD, Stiegler GL, Hallick RB: Detection of multiple, unspliced precursor mRNA transcripts for the 32,000 Mr thylakoid membrane protein from Euglena gracilis chloroplasts. Nucl Acids Res 12: 2001–2017 (1984).
Katoh S, San Pietro A: Ascorbate-supported NADP photoreduction by heated Euglena chloroplasts. Arch Biochem Biophys 122: 144–152 (1967).
Keller M, Weil JH, Nair CKK: Nucleotide sequence of the psbB gene of Euglena gracilis. Plant Molec Biol 13: 723–725 (1989).
Klein RR, Mullet JE: Light-induced transcription of chloroplast genes. psbA transcription is differentially enhanced in illuminated barley. J Biol Chem 265: 1895–1902 (1990).
Kulkarni RD, Mueller UW, Goden SS: Nucleotide sequence of psbB from Synechococcus sp. strain PCC7942. Biochim Biophys Acta 1173: 329–332 (1993).
Kusnetsov VV, Mikulovich TP, Kukina IM, Cherepneva GN, Herrmann RG, Kulaeva ON: Changes in the level of chloroplast transcripts in pumpkin cotyledons during heat shock. FEBS Lett 321: 189–193 (1993).
Mamedov M, Hayashi H, Murata N: Effects of glycine-betaine and unsaturation of membrane lipids on heat stability of photosynthetic electron transport and phosphorylation reactions in Synechocystis PCC6803. Biochim Biophys Acta 1142: 1–5 (1993).
Manzara T, Hu JX, Hallick RB: Characterization of the trnD, trnK, psaA locus of Euglena gracilis chloroplast DNA. Plant Mol Biol 8: 327–336 (1987).
Ortiz W: Contributions from the plastid and the nucleo-cytoplasmic compartments to the eyents associated with heat-bleaching in Euglena gracilis. J Plant Physiol 139: 543–548 (1992).
Ortiz W, Kutner MS: Cycloheximide delays chlorophyll loss but not the inhibition of plastid protein synthesis during heat-bleaching in Euglena gracilis. J Plant Physiol 137: 110–115 (1990).
Ortiz W, Kutner MS: Factors that affect the synthesis and accumulation of a polypeptide of the light-haryesting chlorophyll-protein complex II in Euglena gracilis. J Plant Physiol 134: 75–80 (1989).
Ortiz W, Reardon EM, Price CA: Preparation of chloroplasts from Euglena highly active in protein synthesis. Plant Physiol 66: 291–294 (1980).
Ortiz W, Wilson CJ: Induced changes in chloroplast protein accumulation during heat-bleaching in Euglena gracilis. Plant Physiol 86: 554–561 (1988).
Pringsheim EG, Pringsheim O: Experimental elimination of chromatophores and eye-spot in Euglena gracilis. New Phytol 51: 65–76 (1952).
Reinbothe S, Reinbothe C, Heintzen C, Seidenbecher C, Parthier B: A methyl jasmonate-induced shift in the length of the 5′ untranslated region impairs translation of the plastid rbcL transcript in barley. EMBO J 12: 1505–1512 (1993).
Schantz R: Mapping of the chloroplast genes coding for the chlorophyll a-binding proteins of E. gracilis. Plant Sci 40: 43–49 (1985).
Schreiber U, Berry JA: Heat-induced changes of chlorophyll fluorescence in intact leaves correlated with damage of the photosynthetic apparatus. Planta 136: 233–238 (1977).
Southern EM: Detection of specific sequences among DNA fragments by gel electrophoresis. J Mol Biol 98: 503–517 (1975).
Uzzo A, Lyman H: The nature of the chloroplast genome of Euglena gracilis. In: Forti G, Avron M, Melandri A (eds) Proceedings Second International Congress on Photosynthesis Research, pp. 2585–2599. Junk, The Hague (1971).
Vermaas WFJ, Ikeuchi M, Inoue Y: Protein composition of the photosystem II core complex in genetically engineered mutants of the cyanobacterium Synechocystis sp. PCC6803. Photosynth Res 17: 97–113 (1988).
Vermaas WFJ, Williams JGK, Rutherford AW, Mathis P, Arntzen CJ: Genetically engineered mutant of the cyanobacterium Synechocystis 6803 lacks the photosystem II chlorophyll binding protein CP47. Proc Natl Acad Sci USA 83: 9474–9477 (1986).
Vierling E, Key JL: Ribulose-1,5-bisphosphate carboxylase synthesis during heat shock. Plant Physiol 78: 155–162 (1985).
Winter U, Feierabend J: Multiple coordinate controls contribute to a balanced expression of ribulose-1,5-bisphosphate carboxylase subunits in rye leaves. Eur J Biochem 187: 445–453 (1990).
Yepiz-Plascencia GM, Radebaugh CA, Hallick RB: The Euglena chloroplast rpoB gene. Novel gene organization and transcription of the RNA polymerase subunit operon. Nucl Acids Res 18: 1869–1878 (1990).
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Thomas, E.J., Ortiz, W. Loss of chloroplast transcripts for proteins associated with photosystem II: an early event during heat-bleaching in Euglena gracilis . Plant Mol Biol 27, 317–325 (1995). https://doi.org/10.1007/BF00020186
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DOI: https://doi.org/10.1007/BF00020186