Skip to main content
SpringerLink
Log in

Photosystem II chlorophyll a fluorescence lifetimes and intensity are independent of the antenna size differences between barley wild-type and chlorina mutants: Photochemical quenching and xanthophyll cycle-dependent nonphotochemical quenching of fluorescence

  • Regular Paper
  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Photosystem II (PS II) chlorophyll (Chl) a fluorescence lifetimes were measured in thylakoids and leaves of barley wild-type and chlorina f104 and f2 mutants to determine the effects of the PS II Chl a+b antenna size on the deexcitation of absorbed light energy. These barley chlorina mutants have drastically reduced levels of PS II light-harvesting Chls and pigment-proteins when compared to wild-type plants. However, the mutant and wild-type PS II Chl a fluorescence lifetimes and intensity parameters were remarkably similar and thus independent of the PS II light-harvesting antenna size for both maximal (at minimum Chl fluorescence level, Fo) and minimal rates of PS II photochemistry (at maximum Chl fluorescence level, Fm). Further, the fluorescence lifetimes and intensity parameters, as affected by the trans-thylakoid membrane pH gradient (ΔpH) and the carotenoid pigments of the xanthophyll cycle, were also similar and independent of the antenna size differences. In the presence of a ΔpH, the xanthophyll cycle-dependent processes increased the fractional intensity of a Chl a fluorescence lifetime distribution centered around 0.4–0.5 ns, at the expense of a 1.6 ns lifetime distribution (see Gilmore et al. (1995) Proc Natl Acad Sci USA 92: 2273–2277). When the zeaxanthin and antheraxanthin concentrations were measured relative to the number of PS II reaction center units, the ratios of fluorescence quenching to [xanthophyll] were similar between the wild-type and chlorina f104. However, the chlorina f104, compared to the wild-type, required around 2.5 times higher concentrations of these xanthophylls relative to Chl a+b to obtain the same levels of xanthophyll cycle-dependent fluorescence quenching. We thus suggest that, at a constant ΔpH, the fraction of the short lifetime distribution is determined by the concentration and thus binding frequency of the xanthophylls in the PS II inner antenna. The ΔpH also affected both the widths and centers of the lifetime distributions independent of the xanthophyll cycle. We suggest that the combined effects of the xanthophyll cycle and ΔpH cause major conformational changes in the pigment-protein complexes of the PS II inner or core antennae that switch a normal PS II unit to an increased rate constant of heat dissipation. We discuss a model of the PS II photochemical apparatus where PS II photochemistry and xanthophyll cycle-dependent energy dissipation are independent of the Peripheral antenna size.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

Ax:

antheraxanthin

BSA:

bovine serum albumin

cx :

lifetime center of fluorescence decay component x

CP:

chlorophyll binding protein of PS II inner antenna

DCMU:

3-(3,4-dichlorophenyl)-1,1-dimethylurea

DTT:

dithiothreitol

fx :

fractional intensity of fluorescence lifetime component x

Fm, Fm′ :

maximal PS II Chl a fluorescence intensity with all QA reduced in the absence, presence of thylakoid membrane energization

Fo :

minimal PS II Chl a fluorescence intensity with all QA oxidized

Fv=Fm−Fo :

variable level of PS II Chl a fluorescence

HPLC:

high performance liquid chromatography

kA :

rate constant of all combined energy dissipation pathways in PS II except photochemistry and fluorescence

kF :

rate constant of PS II Chl a fluorescence

LHCIIb:

main light harvesting pigment-protein complex (of PS II)

Npig :

mols Chl a+b per PS II

NPQ=(Fm/Fm′−1):

nonphotochemical quenching of PS II Chl a fluorescence

PAM:

pulse-amplitude modulation fluorometer

PFD:

photon-flux density, μmols photons m−2 s−1

PS II:

Photosystem II

P680:

special-pair Chls of PS II reaction center

QA :

primary quinone electron acceptor of PS II

Vx :

violaxanthin

wx :

width at half maximum of Lorentzian fluorescence lifetime distribution x

Zx:

zeaxanthin

ΔpH:

trans-thylakoid proton gradient

% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% GabaqaaiGacaGaamqadaabaeaafiaakeaacqGH8aapcqaHepaDcqGH% +aGpdaWgaaWcbaGaamOraiaad2gaaeqaaaaa!4989!\[< \tau > _{Fm}\],% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% GabaqaaiGacaGaamqadaabaeaafiaakeaacqGH8aapcqaHepaDcqGH% +aGpdaWgaaWcbaGaamOraiaad+gaaeqaaOGaeyypa0Zaaabqaeaaca% WGMbWaaSbaaSqaaiaadIhaaeqaaOGaam4yamaaBaaaleaacaWG4baa% beaaaeqabeqdcqGHris5aaaa!50D3!\[< \tau > _{Fo} = \sum {f_x c_x }\]:

average lifetime of Chl a fluorescence calculated from a multi-exponential model under Fm, Fo conditions

References

  • Adams WWIII, Demmig-Adams B and Winter K (1990a) Relative contributions of zeaxanthin-related and zeaxanthin unrelated types of ‘high-energy-state’ quenching of chlorophyll fluorescence in spinach leaves exposed to various environmental conditions. Plant Physiol 92: 302–309

    Google Scholar 

  • Adams WWIII, Demmig-Adams B, Winter K and Schreiber U (1990b) The ratio of variable to maximum fluorescence from Photosystem II, measured in leaves at ambient temperature and at 77K, as an indicator of the photon yield of photosynthesis. Planta 180: 166–174

    Article  Google Scholar 

  • Andrews JR, Fryer MJ and Baker NR (1995) Consequences of LHCII deficiency for photosynthetic regulation in chlorina mutants of barley. Photosynth Res 44: 81–91

    Google Scholar 

  • Bassi R, Hinz U and Barbato R (1985) The role of light harvesting complex and Photosystem II in thylakoid stacking in the chlorina-f2 barley mutant. Carlsberg Res Commun 50: 347–367

    Google Scholar 

  • Bassi R, Pineau B, Dainese P and Marquardt J (1993) Carotenoid-binding proteins of Photosystem II. Eur J Biochem 212: 297–303

    PubMed  Google Scholar 

  • Bilger W and Björkman O (1991) Temperature dependence of violaxanthin de-epoxidation and non-photochemical fluorescence quenching in intact leaves of Gossypium hirsutum L. and Malva parviflora L. Planta 184: 226–234

    Article  Google Scholar 

  • Björkman O and Demmig B (1987) Photon yield of O2 evolution and chlorophyll fluorescence characteristics at 77K among vascular plants of diverse origins. Planta 170: 489–504

    Article  Google Scholar 

  • Briantais J-M (1994) Light-harvesting chlorophyll a−b complex requirement for regulation of Photosystem II photochemistry by non-photochemical quenching. Photosynth Res 40: 287–294

    Article  Google Scholar 

  • Briantais J-M, Vernotte C, Krause GH and Weis E (1986) Chlorophyll a fluorescence of higher plants: Chloroplasts and leaves. In: Govindjee, Amesz J and Fork DC (eds) Light Emission by Plants and Bacteria, pp 539–586. Academic Publishers, Orlando, FL

    Google Scholar 

  • Briantais J-M, Dacosta J, Goulas Y, Ducruet J-M and Moya I (1996) Heat stress induces in leaves an increase of the minimum level of chlorophyll fluorescence, Fo. A time-resolved analysis. Photosynth Res 48: 189–196 (this issue)

    Google Scholar 

  • Butler W and Strasser RJ (1977) Tripartite model for the photochemical apparatus of green plant photosynthesis. Proc Nat Acad Sci USA 74: 3382–3385

    PubMed  Google Scholar 

  • Crofts AR and Yerkes CT (1994) A molecular mechanism for qE-quenching. FEBS Lett 352: 265–270

    Article  Google Scholar 

  • Dau H (1994) Molecular mechanisms and quantitative models of variable Photosystem II fluorescence. Photochem Photobiol 60: 1–23

    Google Scholar 

  • Dau H and Sauer K (1996) Exciton equilibration and Photosystem II exciton dynamics-a fluorescence study on Photosystem II membrane particles of spinach. Biochim Biophys Acta 1273: 175–190

    Google Scholar 

  • Demmig-Adams B, Gilmore AM and Adams WWIII (1996) In vivo functions of carotenoids in higher plants. FASEB 10: 203–214

    Google Scholar 

  • Emerson R and Arnold A (1932a) A separation of the reactions in photosynthesis by means of intermittent light. J Gen Physiol 15: 391–420

    Article  Google Scholar 

  • Emerson R and Arnold A (1932b) The photochemical reaction in photosynthesis. J Gen Physiol 16: 191–205

    Article  Google Scholar 

  • Falbel TG, Staehelin A and Adams WWIII (1994) Analysis of xanthophyll cycle carotenoids and chlorophyll fluorescence in light intensity-dependent chlorophyll-deficient mutants of wheat and barley. Photosynth Res 42: 191–202

    Google Scholar 

  • Falk S, Bruce D and Huner NPA (1994a) Photosynthetic performance and fluorescence in relation to antenna size and absorption cross-sections in rye and barley grown under normal and intermittent light conditions. Photosynth Res 42: 145–155

    Google Scholar 

  • Falk S, Król M, Maxwell DP, Rezansoff DA, Gray GR and Huner NPA. (1994b) Changes in the in vivo fluorescence quenching in rye and barley as a function of reduced PS II light-harvesting antenna size. Physiol Plant 91: 551–558

    Article  Google Scholar 

  • Frauenfelder H, Parak F and Young RD (1988) Conformational substates in proteins. Ann Rev Biophys Biophys Chem 17: 451–479

    Article  Google Scholar 

  • Gaffron H and Wohl K (1936) Zur Theorie der Assimilation. Naturwissenschaften 24: 81–90, 103–107

    Google Scholar 

  • Gilmore AM and Björkman O (1995) Temperature-sensitive coupling and uncoupling of ATPase-mediated, nonradiative energy dissipation: Similarities between chloroplasts and leaves. Planta 197: 646–654

    Article  Google Scholar 

  • Gilmore AM and Yamamoto HY (1991) Resolution of lutein and zeaxanthin using a nonendcapped, lightly carbon-loaded C-18 high-performance liquid chromatographic column. J Chromatogr 543: 137–145

    Article  Google Scholar 

  • Gilmore AM and Yamamoto HY (1992) Dark induction of zeaxanthin-dependent nonphotochemical fluorescence quenching mediated by ATP. Proc Natl Acad Sci USA 89: 1899–1903

    PubMed  Google Scholar 

  • Gilmore AM and Yamamoto HY (1993) Linear models relating xanthophylls and lumen acidity to non-photochemical fluorescence quenching. Evidence that antheraxanthin explains zeaxanthin-independent quenching. Photosynth Res 35: 67–78

    Google Scholar 

  • Gilmore AM, Hazlett TL and Govindjee (1995a) Xanthophyll cycle dependent quenching of Photosystem II chlorophyll a fluorescence: formation of a quenching complex with a short fluorescence lifetime. Proc Natl Acad Sci USA 92: 2273–2277

    PubMed  Google Scholar 

  • Gilmore AM, Hazlett TL, Björkman O and Govindjee (1995b) Xanthophyll cycle dependent non-photochemical quenching of chlorophyll a fluorescence at low physiological temperatures. In: Mathis P (ed) Photosynthesis: From Light to Biosphere, Vol IV, pp 825–828. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Govindjee (1995) Sixty-three years since Kautsky: Chlorophyll a fluorescence. Aust J Plant Physiol 22: 131–160

    Google Scholar 

  • Govindjee, Van de Ven M, Cao J, Royer C and Gratton E. (1993) Multifrequency cross-correlation phase fluorometry of chlorophyll a fluorescence in thylakoid and PS II-enriched membranes. Photochem Photobiol 58: 438–445

    PubMed  Google Scholar 

  • Graan T and Ort DR (1984) Quantitation of the rapid electron donors to P700, the functional plastoquinone pool, and the ratio of the photosystems in spinach chloroplasts. J Biol Chem 22: 14003–14010

    Google Scholar 

  • Hager A (1969) Lichtbedingte pH-Erniedrigung in einem Chloroplasten-Kompartiment als Ursache der enzymatischen Violaxanthin→Zeazanthin-Umwandlung; Beziehungen zur Photophosphorylierung. Planta 89: 224–243

    Google Scholar 

  • Harrison MA, Nemson JA and Melis A (1993) Assembly and composition of the chlorophyll a−b light-harvesting complex of barley (Hordeum vulgare L.): Immunochemical analysis of chlorophyll b-less and chlorophyll b-deficient mutants. Photosynth Res 38: 141–151

    Google Scholar 

  • Härtel H and Lokstein H (1995) Relationship between quenching of maximum and dark-level chlorophyll fluorescence in vivo: dependence on Photosystem II antenna size. Biochim Biophys Acta 1228: 91–94

    Google Scholar 

  • Holzwarth AR (1988) Time resolved chlorophyll fluorescence. What kind of information does it provide? In: Lichtenthaler HK (ed) Applications of Chlorophyll Fluorescence in Photosynthesis, Stress Physiology, Hydrobiology and Remote Sensing, pp 21–31. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Horton P, Ruban AV and Walters RG (1994) Regulation of light harvesting in green plants. Plant Physiol 106: 415–420

    PubMed  Google Scholar 

  • Jahns P and Schweig S (1995) Energy-dependent fluorescence quenching in thylakoids from intermittent light grown pea plants-evidence for an interaction of zeaxanthin and the chlorophyll a/b binding protein CP26. Plant Physiol Biochem 33: 683–687

    Google Scholar 

  • Karukstis KK and Sauer K (1984) Organization of the photosynthetic apparatus of the chlorina-f2 mutant of barley using chlorophyll fluorescence decay kinetics. Biochim Biophys Acta 766: 148–155

    PubMed  Google Scholar 

  • Knoetzel J and Simpson D (1991) Expression and organization of antenna proteins in the light- and temperature-sensitive barley mutant chlorina-104. Planta 185: 111–123

    Article  Google Scholar 

  • Knox RS (1975) Excitation energy transfer and migration: Theoretical considerations. In: Govindjee (ed) Bioenergetics of Photosynthesis, pp 183–224. Academic Press, New York

    Google Scholar 

  • Knox RS and Lin S (1988). Time resolution and kinetics of ‘F680’ at low temperatures in spinach chloroplasts. In: Scheer H and Schneider S (eds) Photosynthetic Light Harvesting Systems: Structure and Function, pp 567–577. Walter de Gruyter, Berlin

    Google Scholar 

  • Krugh BW and Miles D (1995) Energy transfer for low temperature fluorescence in PS II mutant thylakoids. Photosynth Res 44: 117–125

    Google Scholar 

  • Leverenz JW, Öquist G and Wingsle G (1992) Photosynthesis and photoinhibition in leaves of chlorophyll b-less barley in relation to absorbed light. Physiol Plant 85: 495–502

    Article  Google Scholar 

  • Lin S and Knox RS (1991) Studies of excitation energy transfer within the green alga Chlamydomonas reinhardtii and its mutants at 77K. Photosynth Res 27:157–168

    Google Scholar 

  • Lokstein H, Härtel H, Hoffman P, Woitke P and Renger G (1994) The role of light-harvesting complex II in excess excitation energy dissipation: An in-vivo fluorescence study on the origin of high-energy quenching. Photochem Photobiol 26: 175–184

    Article  Google Scholar 

  • Mills JD and Mitchell P (1982) Modulation of coupling factor ATPase activity in intact chloroplasts. Reversal of thiol modulation in the dark. Biochim Biophys Acta 679: 75–83

    Google Scholar 

  • Pearlstein RM (1982) Chlorophyll singlet excitons. In: Govindjee (ed) Photosynthesis: Energy Conversion by Plants and Bacteria, pp 294–391. Academic Press, New York

    Google Scholar 

  • Petrack B and Lipman F. (1961) Photophosphorylation and photohydrolysis in cell-free preparations of blue-green algae. In: McElroy WD and Glass B (eds) Light and Life, pp 621–630. Johns Hopkins Press, Baltimore, MD

    Google Scholar 

  • Porra RJ, Thompson WA and Kriedmann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: Verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975: 384–394

    Google Scholar 

  • Preiss S and Thornber JP (1995) Stability of the apoproteins of light-harvesting complex I and II during biogenesis of thylakoids in the chlorophyll b-less barley mutant chlorina f-2. Plant Physiol 107: 709–717

    PubMed  Google Scholar 

  • Rijgersberg CP, Amesz J, Thielen APGM and Swager JA (1979) Fluorescence emission spectra of chloroplasts and subchloroplast preparations at low temperatures. Biochim Biophys Acta 545: 473–482

    PubMed  Google Scholar 

  • Schatz GH, Brock H and Holzwarth AR (1988) A kinetic and energetic model for the primary processes in Photosystem II. Biophys J 54: 397–405

    Google Scholar 

  • Searle GFW, Tredwell CJ, Barber J and Porter G (1979) Picosecond time-resolved study of chlorophyll organization and excitation energy distribution in chloroplasts from wild-type barley and a mutant lacking chlorophyll b. Biochim Biophys Acta 545: 496–507

    PubMed  Google Scholar 

  • Simpson D, Machold O, Høyer-Hansen G and von Wettstein D (1985) Chlorina mutants of barley (Hordeum vulgare L.). Carlsberg Res Commun 50: 223–238

    Google Scholar 

  • Tyystjärvi E, Kettunen R and Aro E-M (1994) The rate constant of photoinhibition in vitro is independent of the antenna size of Photosystem II but depends on temperature. Biochim Biophys Acta 1186: 177–185

    Google Scholar 

  • van Grondelle R, Dekker JP, Gillbro T and Sundstrom V (1994) Energy transfer and trapping in photosynthesis. Biochim Biophys Acta 1187: 1–65

    Google Scholar 

  • van Kooten O and Snel JFH (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth Res 25: 147–150

    Google Scholar 

  • Visser HM, Somsen OJG, van Mourik F, Lin S, van Stokkum IHM and van Grondelle R (1995) Direct observation of sub-picosecond equilibration of excitation energy in the light-harvesting antenna of Rhodospirillum rubrum. Biophys J 69: 1083–1099

    PubMed  Google Scholar 

  • Walters RG, Ruban AV and Horton P (1994) Higher plant light-harvesting complexes LHCIIa and LHCIIc are bound by dicyclohexylcarbodiimide during inhibition of energy dissipation. Eur J Biochem 226: 1063–1069

    PubMed  Google Scholar 

  • Weis E and Berry JA (1987) Quantum efficiency of Photosystem II in relation to ‘energy’-dependent quenching of chlorophyll fluorescence. Biochim Biophys Acta 894: 198–207

    Google Scholar 

  • Yamamoto HY and Bassi R (1996) Carotenoids: Localization and Function. In: Ort DR and Yocum CF (eds) Oxygenic Photosynthesis: The Light Reactions. Advances in Photosynthesis, Vol 4, pp 539–563. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Yamamoto HY and Kamite L (1972) The effects of dithiothreitol on violaxanthin de-epoxidation and absorbance changes in the 500-nm region. Biochim Biophys Acta 267: 538–543

    PubMed  Google Scholar 

  • Yamamoto HY, Nakayama TOM and Chichester CO (1962) Studies on the light and dark interconversions of leaf xanthophylls. Arch Biochem Biophys 97: 168–173

    PubMed  Google Scholar 

Download references

Author information

Author notes

  1. Adam M. Gilmore

    Present address: ANU/RSBS Photobioenergetics Group, 0200, Canberra, ACT, Australia

Authors and Affiliations

  1. Department of Plant Biology, University of Illinois at Urbana-Champaign, 61801-3707, Urbana, IL, USA

    Adam M. Gilmore & Govindjee

  2. Laboratory for Fluorescence Dynamics, University of Illinois at Urbana-Champaign, 61801-3707, Urbana, IL, USA

    Theodore L. Hazlett

  3. Department of Physics, University of Illinois at Urbana-Champaign, 61801-3707, Urbana, IL, USA

    Peter G. Debrunner

Authors
  1. Adam M. Gilmore
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Theodore L. Hazlett
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter G. Debrunner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Govindjee
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gilmore, A.M., Hazlett, T.L., Debrunner, P.G. et al. Photosystem II chlorophyll a fluorescence lifetimes and intensity are independent of the antenna size differences between barley wild-type and chlorina mutants: Photochemical quenching and xanthophyll cycle-dependent nonphotochemical quenching of fluorescence. Photosynth Res 48, 171–187 (1996). https://doi.org/10.1007/BF00041007

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00041007

Key words

Search

Navigation