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Characterization of a novel drought-induced 34-kDa protein located in the thylakoids ofSolanum tuberosum L. plants

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Abstract

Using two-dimensional electrophoresis and Coomassie Blue staining, the accumulation of a 34-kDa protein (named cdsp 34 for chloroplastic drought-induced stress protein) is shown in the thylakoids ofSolanum tuberosum plants subjected to a progressive and reversible water deficit. In-vivo labeling experiments showed an increased synthesis of cdsp 34 from the early stages of drought stress (leaf relative water content around 85%) and throughout the constraint. Sequences of the N-terminal part and of four tryptic-digest peptides did not reveal significant homology between the cdsp 34 protein and other known proteins. Western blotting analysis, using a serum raised against the N-terminal part of cdsp 34, confirmed the accumulation of cdsp 34 in thylakoids upon drought stress. From immunoblot analysis of different chloroplastic subfractions, the cdsp 34 protein appears to be an extrinsic protein preferentially located in unstacked stroma thylakoids. Immunoprecipitation of in-vitro-translated products, as well as Southern analysis, showed that the cdsp 34 protein is nuclear encoded. After rewatering of water-stressed plants, the level of cdsp 34 synthesis was reduced, but remained substantially higher than in control plants. Western analysis showed the persistence of a high amount of cdsp 34 in rewatered plants for at least two weeks. Based on the abundance and on the location of cdsp 34 within thylakoids, a putative role for this novel chloroplastic protein is discussed in relation to the tolerance of the photosynthetic apparatus of higher plants to dehydration.

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Abbreviations

2-D:

two-dimensional

CF:

coupling factor

cdsp:

chloroplastic drought-induced stress protein

dsp:

desiccation stress protein

LHC:

light-harvesting complex

PVDF:

polyvinylidene fluoride

RWC:

relative water content

References

  • Adamska I, Ohad I, Kloppstech K (1992) Synthesis of the early light-inducible protein is controlled by blue light and related to light stress. Proc Natl Acad Sci USA 89: 2610–2613

    Google Scholar 

  • Baker JC, Steele C, Dure III L (1988) Sequence and characterization of 6 Lea proteins and their genes from cotton. Plant Mol Biol 11: 277–291

    Google Scholar 

  • Bartels D, Hanke C, Schneider K, Michel D, Salamini F (1992) A desiccation-related Elip-like gene from the resurrection plant Craterostigma plantagineum is regulated by light and ABA. EMBO J 11: 2771–2778

    Google Scholar 

  • Bartels D, Alexander R, Schneider K, Elster R, Velasco R, Alamillo J, Bianchi G, Nelson D, Salamini F (1993) Desiccation-related gene products analyzed in a resurrection plant and in barley embryos. Current topics in plant physiology, an american society of plant physiologists series, vol 10, Rockville MD, USA, pp 119–127

  • Bergman A, Gardeström P, Ericson I (1980) Method to obtain a chlorophyll-free preparation of intact mitochondria from spinach leaves. Plant Physiol 66: 442–445

    Google Scholar 

  • Bray E (1990) Drought-stress-induced polypeptide accumulation in tomato leaves. Plant Cell Environ 13: 531–538

    Google Scholar 

  • Bray E (1993) Molecular responses to water deficit. Plant Physiol 103:1035–1040

    Google Scholar 

  • Cai D, Herrmann RG, Klösgen RB (1993) The 20 kDa apoprotein of the CP24 complex of photosystem II: an alternative model to study import and intra-organellar routing of nuclear-encoded thylakoid proteins. Plant J 3: 383–392

    Google Scholar 

  • Chaves MM (1991) Effects of water deficits on carbon assimilation. J Exp Bot 234: 1–16

    Google Scholar 

  • Conroy JP, Virgona JM, Smillie RM, Barlow EW (1988) Influence of drought acclimation and C02 enrichment on osmotic adjustment and chlorophyll a fluorescence of sunflower during drought. Plant Physiol 86: 1108–1115

    Google Scholar 

  • Comic G, Ghashghaie J, Genty B, Briantais JM (1992) Leaf photosynthesis is resistant to mild drought stress. Photosynthetica 27: 295–309

    Google Scholar 

  • Damerval C, De Vienne D, Zivy M, Thiellement H (1986) Technical improvement in two-dimensional electrophoresis. Increased level of genetic variation detected in wheat seedling proteins. Electrophoresis 7: 52–54

    Google Scholar 

  • Dure III L (1993) Structural motifs in LEA proteins. In: Close TJ, Bray EA (eds) Plant responses to cellular dehydration during environmental stress. Current topics in plant physiology, an american society of plant physiologists series, vol 10, Rockville MD, USA, pp 91–103

  • Gatenby AA, Castleton JA, Saul MW (1981) Expression inE. coli of maize and wheat chloroplast genes for large subunit of ribulose bisphosphate carboxylase. Nature 291: 117–121

    Google Scholar 

  • Guillemault P, Maréchal-Drouart L (1992) Isolation of plant DNA: a fast, inexpensive and reliable method. Plant Mol Biol Rep 10: 60–65

    Google Scholar 

  • Guy CL, Niemi KJ, Brambl R (1985) Altered gene expression during cold acclimation of spinach. Proc Natl Acad Sci USA 82: 3673–3677

    Google Scholar 

  • Havaux M (1992) Stress tolerance of photosystem II in vivo. Antagonistic effects of water, heat, and photoinhibition stresses. Plant Physiol 100: 424–432

    Google Scholar 

  • Kaiser WM (1987) Effects of water deficit on photosynthetic capacity. Physiol Plant 71: 142–149

    Google Scholar 

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

    Google Scholar 

  • Kloppstech K, Meyer G, Schuster G, Ohad I (1985) Synthesis, transport and localization of a nuclear 22-kD heat-shock protein in the chloroplast membranes of peas andChlamydomonas reinhardi. EMBO J 4:1901–1909

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  • Lichtenthaler HK, Wellburn AR (1983) Determination of total carotenoids and chlorophyllsa andb of leaf extracts in different solvants. Biochem Soc Trans 603: 591–592

    Google Scholar 

  • Lipman DJ, Pearson W (1985) Rapid and sensitive protein similarity searches. Science 227: 1435–1441

    Google Scholar 

  • Lunn JE, Droux M, Martin J, Douce R (1990) Localization of ATP sulfurylase andO-acetylserine(thiol)lyase in spinach leaves. Plant Physiol 94: 1345–1352

    Google Scholar 

  • Maslenkova LT, Miteva TS, Popova LP (1992) Changes in the polypeptide patterns of barley seedlings exposed to jasmonic acid and salinity. Plant Physiol 98: 700–707

    Google Scholar 

  • Meyer S, Hung SPN, Trémolières A, de Kouchkovsky Y (1992) Energy coupling, membrane lipids and structure of thylakoids of lupin plants submitted to water stress. Photosynth Res 32: 95–107

    Google Scholar 

  • Mills WR, Joy KW (1980) A rapid method for isolation of purified physiologically active chloroplasts used to study the intracellular distribution of amino acids in pea leaves. Planta 148: 75–83

    Google Scholar 

  • O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021

    Google Scholar 

  • Oishi K, Sumnicht T, Tewari KK (1981) Messenger ribonucleic acid transcripts of pea chloroplast deoxyribonucleic acid. Biochemistry 20: 5710–5717

    Google Scholar 

  • Roscoe TJ, Ellis RJ (1982) Two-dimensional gel electrophoresis of chloroplast proteins. In: Edelman M, Hallick RB, Chua NH (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, Amsterdam, The Netherlands, pp 1015–1028

    Google Scholar 

  • Sane PV, Goodchild DJ, Park PB (1970) Characterization of chloroplast PS I and PS II separated by a nondetergent method. Biochem Biophys Acta 216: 162–178

    Google Scholar 

  • Santarius KA (1973) The protective effects of sugars on chloroplast membranes during temperature and water stress and its relationship to frost, desiccation and heat resistance. Planta 113: 105–114

    Google Scholar 

  • Santarius KA, Müller M (1979) Investigations on heat resistance of spinach leaves. Planta 146: 529–538

    Google Scholar 

  • Schneider K, Wells B, Schmelzer E, Salamini F, Bartels D (1993) Desiccation leads to the rapid accumulation of both cytosolic and chloroplastic proteins in the resurrection plantCraterostigma plantagineum Hochst. Planta 189: 120–131

    Google Scholar 

  • Schuster G, Even D, Kloppstech K, Ohad I (1988) Evidence for protection by heat-shock proteins against photoinhibition during heat-shock. EMBO J 7: 1–6

    Google Scholar 

  • Seemann JR, Downtown WD, Berry JA (1986) Temperature and leaf osmotic potential as factors in the acclimation of photosynthesis to high temperature in desert plants. Plant Physiol 80: 926–930

    Google Scholar 

  • Skriver K, Mundy J (1990) Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512

    Google Scholar 

  • Steinback KE, Mullet JE, Arntzen CJ (1982) Fractionation of thylakoid membrane protein complexes by sucrose densitygradient centrifugation. In: Edelman M, Hallick RB, Chua NH (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, Amsterdam, The Netherlands, pp 863–872

    Google Scholar 

  • Stone KL, Williams KR (1993) Enzymatic digestion of proteins and HPLC peptide isolation. In: Matsudaira P (ed) A practical guide to protein and peptide purification for microsequencing, second edition. Academic Press Inc, San Diego, Cal., USA, pp 43–69

    Google Scholar 

  • Tourneux C, Peltier G (1995) Effect of water deficit on photosynthetic oxygen exchange measured using1802 and mass spectrometry inSolanum tuberosum L. leaf discs. Planta 195: 570–577

    Google Scholar 

  • Wheeler RM, Mackowiak CL, Sager JC, Knott WM, Hinkle CR (1990) Potato growth and yield using nutrient film technique (NFT). Am Potato J 67: 177–187

    Google Scholar 

  • Yu SG, Stefansson H, Romanowska E, Albertsson PA (1994) Two-dimensional electrophoresis of thylakoid membrane proteins and its application to microsequencing. Photosynth Res 41: 475–486

    Google Scholar 

  • Zeevaart JAD, Creelman RA (1988) Metabolism and physiology of abscisic acid. Annu Rev Plant Physiol Mol Biol 39: 439–473

    Google Scholar 

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Correspondence to Pascal Rey.

Additional information

The protein sequence data reported appear in the Swiss-PROT database under the accession number P 80471

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Pruvot, G., Cuiné, S., Peltier, G. et al. Characterization of a novel drought-induced 34-kDa protein located in the thylakoids ofSolanum tuberosum L. plants. Planta 198, 471–479 (1996). https://doi.org/10.1007/BF00620065

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  • DOI: https://doi.org/10.1007/BF00620065

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