Abstract
Bioenergetic properties of thylakoids from plants submitted to a water stress stress (watering stopped for 6–15 days) have been measured in two lupin genotypes characterized as resistant or susceptible to drought. This energy coupling was assessed by flow-force relationships relating the phosphorylation rate to the magnitude of the proton gradient % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% GabaqaaiGacaGaamqadaabaeaafiaakabbaaa6daaahjxzL5gapeqa% aiabgs5aenaaxacabaGaeqiVd0galeqabaGaaiOFaaaakmaaBaaale% aacaWGibWaaWbaaWqabeaacqGHRaWkaaaaleqaaaaa!4D55!\[\Delta \mathop \mu \limits^\~ _{H^ + } \]. The fluorescent probe 9-aminoacridine was used to express, as a ΔpH, the whole % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% GabaqaaiGacaGaamqadaabaeaafiaakabbaaa6daaahjxzL5gapeqa% aiabgs5aenaaxacabaGaeqiVd0galeqabaGaaiOFaaaakmaaBaaale% aacaWGibWaaWbaaWqabeaacqGHRaWkaaaaleqaaaaa!4D55!\[\Delta \mathop \mu \limits^\~ _{H^ + } \] by calibrating fluorescence quenching against the phosphate potential ΔGp in ‘state 4’, i.e., when ATP synthesis is strictly balanced by its hydrolysis. This calibration procedure was shown to be unaffected by treatments. At equal energization (iso-ΔpH), ATP synthesis was halved by a medium stress and disappeared for a more severe stress, whereas ΔpH at equal energy input (light) declined only under a severe drought. For an identical ΔpH, PS 1-driven phosphorylation is always more efficient than PS 2, both in control and stressed plants. Thus, uncoupling is not the cause of the phosphorylation decline; moreover, retention of a ‘micro-chemiosmotic’ type of coupling implies that the distribution of photosystems and ATPases is unchanged. Parallel to these functional alterations, the lipid content of thylakoids dramatically dropped. As galactolipids fell strongly, neutral lipids rose slightly. Fatty acids decreased then increased with stress, yet phosphorylation did not recover in the latter case and membrane permeability to protons remained unaffected. Overall, these observations suggest a preserved thylakoid structure and this was indeed observed on electron micrographs, even for a severe stress. Therefore, the membrane integrity is probably preserved more by the protein network than by the lipid matrix and the loss of the phosphorylating activity mainly reflects a loss of ATPases or at least their inactivation, possibly due to their altered lipid environment. Finally, from the bioenergetic point of view, the susceptible genotype was unexpectedly less affected by drought than the resistant.
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Abbreviations
- ATPase:
-
ATP-synthase (or hydrolase), also called coupling factor, CF0-CF1
- chl:
-
chlorophyll (a+b)
- DAG, TAG:
-
di-, tri-acylglycerol
- % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% GabaqaaiGacaGaamqadaabaeaafiaakabbaaa6daaahjxzL5gapeqa% aiabgs5aenaaxacabaGaeqiVd0galeqabaGaaiOFaaaakmaaBaaale% aacaWGibWaaWbaaWqabeaacqGHRaWkaaaaleqaaaaa!4D55!\[\Delta \mathop \mu \limits^\~ _{H^ + } \], ΔpH, ΔΨ:
-
transmembrane differences of proton electrochemical potential (‘proton gradient’), of pH, of electrical potential
- ΔGp:
-
‘phosphate potential’, i.e., Gibb's free energy (enthalpy) of phosphorylation
- DGDG, MGDG:
-
di-, monogalactosyl-diacylglycerol
- DMQ:
-
2,5-dimethylquinone
- FFA:
-
free fatty acid
- NL:
-
neutral lipid
- PG:
-
phosphatidylglycerol
- % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% GabaqaaiGacaGaamqadaabaeaafiaakabbaaa6daaahjxzL5gapeqa% aiabfI6aznaaBaaaleaacaWG3baabeaaaaa!4986!\[\Psi _w \], % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% GabaqaaiGacaGaamqadaabaeaafiaakabbaaa6daaahjxzL5gapeqa% aiabfI6aznaaBaaaleaacqaHapaCaeqaaaaa!4A47!\[\Psi _\pi \]:
-
water, osmotic potential
- PS 1, PS 2:
-
Photosystem 1, Photosystem 2
- PYO:
-
pyocyanine
- R:
-
resistant plant
- S:
-
susceptible plant
- Ve, Vi :
-
volumes of the suspending medium, of the internal lumen of thylakoids
References
Alieva SA, Tageeva SV, Tairbokov MG, Kasatkina VS and Vagabova ME (1971) Structural and functional condition of the chloroplasts as a function of the water regime. Sov Plant Physiol 18: 416–422 (engl. transl.)
de Bilderling N and Lourtioux A (1976) Quinze années de phytotronique. In: Jacques R (ed) Etudes de biologie végétale, Hommage au professeur Pierre Chouard, pp 331–341. CNRS, Paris
Bizouarn T, de Kouchkovsky Y and Haraux F (1989) Photophosphorylation at variable ADP concentration but constant ΔpH in lettuce thylakoids. Effect of ΔpH and phosphate on the apparent affinity for ADP. Biochim Biophys Acta 974: 104–113
Bizouarn T, Phung Nhu Hung S, Haraux F and de Kouchkovsky Y (1990) Ionic composition of the medium, surface potential and affinity of the membrane-bound chloroplast ATPase for its charged substrate ADP. Bioelectrochem Bioenerg 24: 215–230
Bligh EG and Dyer WS (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917
Casadio R (1991) Measurements of transmembrane pH differences of low extents in bacterial chromatophores. A study with the fluorescent probe 9-amino, 6-chloro, 2-methoxyacridine. Eur J Biophys 19: 189–201
Chetal S, Wagle DS and Nainawatee HS (1981) Glycolipid changes in wheat and barley chloroplast under water stress. Plant Sci Lett 20: 225–230
Chetal S, Wagle DS and Nainawatee HS (1983) Phospholipid changes in wheat and barley chloroplast under water stress. Plant Sci Lett 29: 273–278
Cornic G, Prioul J-L and Louason G (1983) Stomatal and non-stomatal contribution in the decline in leaf net CO2 uptake during rapid water stress. Physiol Plant 58: 295–301
Fellows RJ and Boyer JS (1976) Structure and activity of chloroplasts of sunflower leaves having various water potentials. Planta 132: 229–239
Ferguson SJ (1985) Fully delocalised chemiosmotic or localised proton flow pathways in energy coupling? A scrutinity of experimental evidence. Biochim Biophys Acta 811: 47–95
Ferrari-Iliou R, Pham Thi AT and Vieira da Silva J (1984) Effect of water stress on the lipid and fatty acid composition of cotton (Gossypium hirsutum) chloroplasts. Physiol Plant 62: 219–224
Gräber P, Schlodder E and Witt HT (1977) Conformational change of chloroplast ATPase induced by a transmembrane electric field and its correlation to phosphorylation. Biochim Biophys Acta 461: 426–440
Haraux F and de Kouchkovsky Y (1980) Measurement of chloroplast internal protons with 9-aminoacridine. Probe binding, dark proton gradient, and salt effects. Biochim Biophys Acta 592: 153–168
Haraux F and de Kouchkovsky Y (1983) The energy transduction theories: A microchemiosmotic approach in thylakoids. Physiol Vég 21: 563–576
Haraux F, Sigalat C, Moreau A and de Kouchkovsky Y (1983) The efficiency of energized protons for ATP synthesis depends on the membrane topography in thylakoids. FEBS Lett 155: 248–252
Hsiao TC (1973) Plant responses to water stress. Annu Rev Plant Physiol 24: 519–570
Hubac C, Guerrier D, Ferran J and Trémolières A (1989) Change of leaf lipid composition during water stress in two genotypes of Lupinus albus L. resistant or susceptible to drought. Plant Physiol Biochem 27: 737–744
Keck RW and Boyer JS (1974) Chloroplast response to low leaf water potentials. III. Differing inhibition of electron transport and photophosphorylation. Plant Physiol 53: 474–479
de Kouchkovsky Y, Haraux F and Sigalat C (1984) A microchemiosmotic interpretation of energy-dependent processes in biomembrane based on the photosynthetic behaviour of thylakoids. Bioelectrochem Bioenerg 13: 143–163
Kurkova EB and Motorina MV (1971) Chloroplast ultrastructure and photosynthesis at different rates of dehydration. Sov Plant Physiol 21: 40–44 (engl. transl.)
Mangold HK (1961) Thin layer chromatography of lipids. J Am Oil Chem Soc 38: 708–727
Mangold HK (1969) Stahl E (ed) Aliphatic Lipids in Thin Layer Chromatography, pp 363–421. Springer-Verlag, New York
Maroti I, Tuba Z and Csik M (1984) Changes of chloroplast ultrastructure and carbohydrate level in Festuca, Achillea and Sedum during drought and after recovery. J Plant Physiol 116: 1–10
Mayoral ML, Atsmon D, Shimsi D and Gromet-Elhanan Z (1981) Effect of water stress on enzyme activities in wheat and related wild species: carboxylase activity, electron transport and photophosphorylation in isolated chloroplasts. Aust J Plant Physiol 8: 385–393
McCarty RE and Portis AJ (1976) A simple quantitative approach to the coupling of photophosphorylation to electron flow in terms of proton fluxes. Biochemistry 15: 5110–5114
Metcalfe LD, Schmitz AA and Pelka JR (1966) Rapid preparation of fatty acid esters from lipids for gas chromatographic analysis. Anal Chem 38: 514–515
Nishimura M, Ito T and Chance B (1962) Studies on bacterial photophosphorylation. III. A sensitive and rapid method of determination of photophosphorylation. Biochim Biophys Acta 59: 177–182
Oleszko S and Moudrianakis EN (1974) The visualisation of the photosynthetic coupling factor in embedded spinach chloroplasts. J Cell Biol 63: 936–948
Pick U (1988) Deactivation of CF0-CF1 ATPsynthase by uncouplers. Biochemistry 27: 8284–8290
Pick U, Gounaris K, Weiss M and Barber J (1985) Tightly bound sulfolipids in chloroplast CF0-CF1. Biochim Biophys Acta 808: 415–420
Rosing J and Slater EC (1972) The value of ΔG0 for the hydrolysis of ATP. Biochim Biophys Acta 267: 275–290
Scholander PF, Hammel HT, Edda D, Bradstreet FD and Hemminssen EA (1965) Sap pressure in vascular plants. Science 148: 339–346
Schuldiner S, Rottenberg H and Avron M (1972) Determination of ΔpH in chloroplasts. II. Fluorescent amines as a probe for the determination of ΔpH in chloroplasts. Eur J Biochem 25: 64–70
Schwab KB and Heber U (1984) Thylakoid membrane stability in drought-tolerant and drought-sensitive plants. Planta 161: 37–45
Sharkey TD and Badger MR (1982) Effects of water stress on photosynthetic electron transport, photophosphorylation, and metabolic levels of Xanthium strumarium mesophyll cells. Planta 156: 199–206
Sigalat C, Haraux F, de Kouchkovsky F, Phung Nhu Hung S and de Kouchkovsky Y (1985) Adjustable microchemiosmotic character of the proton gradient generated by systems I and II for photosynthetic phosphorylation in thylakoids. Biochim Biophys Acta 809: 403–413
Sigalat C, de Kouchkovsky Y, Haraux F and de Kouchkovsky F (1988) Shift from localized to delocalized protonic energy coupling in thylakoids by permeant amines. Biochim Biophys Acta 934: 375–388
Strottmann H and Lohse D (1988) Determination of the H+/ATP ratio of the H+ transport-coupled reversible chloroplast ATPase reaction by equilibrium studies. FEBS Lett 229: 308–312
Trémolières A and Lepage M (1971) Changes in lipid composition during greening of etiolated pea seedlings. Plant Physiol 47: 329–334
Vallon O, Wollman FA and Olive J (1986) Lateral distribution of the main protein complexes of the photosynthetic apparatus in Chlamydomonas reinhardtii and in spinach: an immunocytochemical study using intact thylakoid membranes and a PS II enriched membrane preparation. Photobiochem Photobiophys 12: 203–220
Younis HM, Boyer JS and Govindjee (1979) Conformation and activity of chloroplast coupling factor exposed to low chemical potential of water in cells. Biochim Biophys Acta 548: 328–340
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Deceased 22 May, 1991; dedicated to her memory.
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Meyer, S., Phung Nhu Hung, S., Trémolières, A. et al. Energy coupling, membrane lipids and structure of thylakoids of Lupin plants submitted to water stress. Photosynth Res 32, 95–107 (1992). https://doi.org/10.1007/BF00035944
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DOI: https://doi.org/10.1007/BF00035944