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Rapidly reversible chlorophyll fluorescence quenching induced by pulses of supersaturating light in vivo

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Abstract

The saturation pulse method provides a means to distinguish between photochemical and non-photochemical quenching, based on the assumption that the former is suppressed by a saturating pulse of light (SP) and that the latter is not affected by the SP. Various types of non-photochemical quenching have been distinguished by their rates of dark relaxation in the time ranges of seconds, minutes, and hours. Here we report on a special type of non-photochemical quenching, which is rapidly induced by a pulse of high-intensity light, when PS II reaction centers are closed, and rapidly relaxes again after the pulse. This high-intensity quenching, HIQ, can be quantified by pulse-amplitude-modulation (PAM) fluorimetry (MULTI-COLOR-PAM, high sensitivity combined with high time resolution) via the quasi-instantaneous post-pulse fluorescence increase that precedes recovery of photochemical quenching in the 100–400-µs range. The HIQ amplitude increases linearly with the effective rate of quantum absorption by photosystem II, reaching about 8% of maximal fluorescence yield. It is not affected by DCMU, is stimulated by anoxic conditions, and is suppressed by energy-dependent non-photochemical quenching (NPQ). The HIQ amplitude is close to proportional to the square of maximal fluorescence yield, Fm′, induced by an SP and varied by NPQ. These properties are in line with the working hypothesis of HIQ being caused by the annihilation of singlet excited chlorophyll a by triplet excited carotenoid. Significant underestimation of maximal fluorescence yield and photosystem II quantum yield in dark-acclimated samples can be avoided by use of moderate SP intensities. In physiologically healthy illuminated samples, NPQ prevents significant lowering of effective photosystem II quantum yield by HIQ, if excessive SP intensities are avoided.

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Abbreviations

AL:

Actinic light

3Car* :

Triplet excited state of carotenoid

Chl:

Chlorophyll a

1Chl* :

Singlet excited state of chlorophyll a

3Chl* :

Triplet excited state of chlorophyll a

DCMU:

3-(3,4-Dichloro-phenyl)-1,1-dimethylurea

rETR:

Relative rate of electron transport estimated from fluorescence parameters

F :

Fluorescence yield of illuminated sample measured briefly before application of SP

F(I):

Fluorescence yield attributed to photosystem I

Fo, Fm:

Minimum and maximum fluorescence yield of dark-adapted sample

Fm′:

Maximum fluorescence yield of illuminated sample

HIQ:

High-intensity quenching of chlorophyll fluorescence yields Fm and Fm′

LED:

Light emitting diode

MC-PAM:

MULTI-COLOR-PAM chlorophyll fluorimeter

ML:

Pulse-modulated fluorescence measuring light

MT:

Multiple-turnover light pulse, the response to which can be measured with high time resolution

NPQ:

Regulated non-photochemical quenching of Chl a fluorescence, calculated as (Fm − Fm′)/Fm′

PAM:

Pulse amplitude modulation

PAR:

Quantum flux density of photosynthetically active radiation

Pheo:

Pheophytin molecules in photosystem II reaction centers

P680:

Reaction center chlorophylls of photosystem II

PS:

Photosystem

SP:

Saturating multiple-turnover light pulse applied for quenching analysis

Y(II):

Effective quantum yield of PS II determined by SP quenching analysis

References

  • Barzda V, Vengris M, Valkunas L, van Grondelle R, van Amerongen H (2000) Generation of fluorescence quenchers from triplet states of chlorophylls in the major light-harvesting complex II from green plants. Biochemistry 39:10468–10477

    Article  CAS  Google Scholar 

  • Breton J, Geacintov NA, Swenberg CE (1979) Quenching of fluorescence by triplet excited states in chloroplasts. Biochim Biophys Acta 548:616–635

    Article  CAS  Google Scholar 

  • Butler WL (1972) On the primary nature of fluorescence yield changes associated with photosynthesis. Proc Natl Acad Sci USA 69:3420–3422

    Article  CAS  Google Scholar 

  • Delosme R (1967) Étude de l’induction de fluorescence des algues vertes et des chloroplastes au début d’une illumination intense. Biochim Biophys Acta 143:108–128

    Article  CAS  Google Scholar 

  • Demmig-Adams B, Garab G. Adams III W, Govindjee (eds) (2014) Non-photochemical quenching and energy dissipation in plants, algae and cyanobacteria. Advances in photosynthesis and respiration, vol. 40, Springer, Dordrecht

    Google Scholar 

  • den Haan GA (1976) Chlorophyll-a fluorescence as a monitor for rapid reactions in system II of photosynthesis. Dissertation, Leiden University

  • Duysens LNM, van der Schatte Olivier TE, den Haan GA (1972) In: Abstr. Int. Congr. on Photobiology, Bochum. Abstr. no. 277

  • Earl HJ, Ennahli S (2004) Estimating photosynthetic electron transport via chlorophyll fluorometry without photosystem II light saturation. Photosynth Res 82:177–186

    Article  CAS  Google Scholar 

  • Franck F, Juneau P, Popovic R (2002) Resolution of the photosystem I and photosystem II contributions to chlorophyll fluorescence of intact leaves at room temperature. Biochim Biophys Acta 1556:239–246

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Genty B, Wonders J, Baker NR (1990) Non-photochemical quenching of Fo in leaves is emission wavelength dependent. Consequences for quenching analysis and its interpretation. Photosynth Res 26:133–139

    Article  CAS  Google Scholar 

  • Gilmore AM, Shinkarev VP, Hazlett TL, Govindjee (1998) Quantitative analysis of the effects of intrathylakoid pH and xanthophyll cycle pigments on chlorophyll a fluorescence lifetime distributions and intensity in thylakoids. Biochemistry 37:13582–13593

    Article  CAS  Google Scholar 

  • Gruber MJ, Chmeliov J, Krüger TPJ, Valkunas L, van Grondelle R (2015) Singlet-triplet annihilation in single LHCII complexes. Phys Chem Chem Phys 17:19844–19853

    Article  Google Scholar 

  • Havurinne V, Mattila H, Antinluoma M, Tyystjärvi E (2018) Unresolved quenching mechanism of chlorophyll fluorescence may invalidate MT saturating pulse analyses of photosynthetic electron transfer in microalgae. Physiol Plant 166:365–379

    Article  Google Scholar 

  • Hormann H, Neubauer C, Schreiber U (1994) On the relationship between chlorophyll fluorescence quenching and the quantum yield of electron transport in isolated thylakoids. Photosynth Res 40:93–106

    Article  CAS  Google Scholar 

  • Klimov VV, Klevanik AV, Shuvalov VA, Krasnovsky AA (1977) Reduction of pheophytin in the primary light reaction of photosystem II. FEBS Lett 82:183–186

    Article  CAS  Google Scholar 

  • Klimov VV, Shuvalov VA, Heber U (1985) Photoreduction of pheophytin as a result of electron donation from the water-splitting system to photosystem-II reaction centers. Biochim Biophys Acta 809:345–350

    Article  CAS  Google Scholar 

  • Klughammer C, Schreiber U (2015) Apparent PS II absorption cross-section and estimation of mean PAR in optically thin and dense suspensions of Chlorella. Photosynth Res 123:77–92

    Article  CAS  Google Scholar 

  • Loriaux SD, Avenson TJ, Welles JM, McDermitt DK, Eckles RD, Riensche B, Genty B (2013) Closing in on maximum yield of chlorophyll fluorescence using a single multiphase flash of sub-saturating intensity. Plant, Cell Environ 36:1755–1770

    Article  CAS  Google Scholar 

  • Markgraf T, Berry J (1990) Measurement of photochemical and non-photochemical quenching: corrections for turnover of PS 2 during steady-state photosynthesis. In: Baltscheffsky M (ed) Current research in photosynthesis, vol IV. Kluwer Academic Publishers, Dordrecht, pp 279–282

    Google Scholar 

  • Neubauer C, Schreiber U (1987) The polyphasic rise of chlorophyll fluorescence upon onset of strong continuous illumination: I. Saturation characteristics and partial control by the photosystem II acceptor side. Z Naturforsch 42c:1246–1254

    Article  Google Scholar 

  • Papageorgiou GC, Govindjee (eds) (2004) Chlorophyll fluorescence: a signature of photosynthesis. Springer, Dordrecht

    Google Scholar 

  • Peterman EJG, Dukker FM, van Grondelle R, van Amerongen H (1995) Chlorophyll a and carotenoid triplet states in light-harvesting complex II of higher plants. Biophys J 69:2670–2678

    Article  CAS  Google Scholar 

  • Pfündel EE (1998) Estimating the contribution of photosystem I to total leaf chlorophyll fluorescence. Photosynth Res 56:185–195

    Article  Google Scholar 

  • Pfündel EE, Klughammer C, Meister A, Cerovic ZG (2013) Deriving fluorometer-specific values of relative PS I fluorescence intensity from quenching of Fo fluorescence in leaves of Arabidopsis thaliana and Zea mays. Photosynth Res 114:189–206

    Article  Google Scholar 

  • Pirson A, Ruppel HG (1962) Über die Induktion einer Teilungshemmung in synchronen Kulturen von Chlorella. Arch Mikrobiol 42:499–505

    Google Scholar 

  • Pospísil P, Skotnica J, Naus J (1998) Low and high temperature dependence of minimum F and maximum F chlorophyll fluorescence in vivo. Biochim Biophys Acta 1363:95–99

    Article  Google Scholar 

  • Santabarbara S, Agostini A, Casazza AP, Zuchelli G, Carbonera D (2015) Carotenoid triplet states in photosystem II: coupling with low-energy states of the core complex. Biochim Biophys Acta 1847:262–275

    Article  CAS  Google Scholar 

  • Schansker G, Tóth SZ, Holzwarth AR (2014) Chlorophyll a fluorescence: beyond the limits of the QA model. Photosynth Res 120:43–58

    Article  CAS  Google Scholar 

  • Schreiber U (2004) Pulse-amplitude (PAM) fluorometry and saturation pulse method. In: Papageorgiou G, Govindjee (eds) Chlorophyll fluorescence: a signature of photosynthesis. Kluwer Academic Publishers, Dordrecht, pp 279–319

    Chapter  Google Scholar 

  • Schreiber U, Neubauer C (1987) The polyphasic rise of chlorophyll fluorescence upon onset of strong continuous illumination: II. Partial control by the photosystem II donor side and possible ways of interpretation. Z Naturforsch 42c:1255–1264

    Article  Google Scholar 

  • Schreiber U, Vidaver W (1974) Chlorophyll fluorescence induction in anaerobic Scenedesmus obliquus. Biochim Biophys Acta 368:97–112

    Article  CAS  Google Scholar 

  • Schreiber U, Vidaver W (1975) Analysis of anaerobic fluorescence decay in Scenedesmus obliquus. Biochim Biophys Acta 387:37–51

    Article  CAS  Google Scholar 

  • Schreiber U, Bilger W, Schliwa U (1986) Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer. Photosynth Res 10:51–62

    Article  CAS  Google Scholar 

  • Schreiber U, Hormann H, Neubauer C, Klughammer C (1995) Assessment of photosystem II photochemical quantum yield by chlorophyll fluorescence quenching analysis. Aust J Plant Physiol 22:209–220

    CAS  Google Scholar 

  • Schreiber U, Klughammer C, Kolbowski J (2012) Assessment of wavelength-dependent parameters of photosynthetic electron transport with a new type of multi-color PAM chlorophyll fluorometer. Photosynth Res 113:127–144

    Article  CAS  Google Scholar 

  • Seaton GGR, Walker DA (1990) Chlorophyll fluorescence as a measure of photosynthetic carbon assimilation. Proc R Soc Lond 242:29–35

    Article  Google Scholar 

  • Siefermann-Harms (1987) The light-harvesting and protective functions of carotenoids in photosynthetic membranes. Physiol Plant 69:561–568

    Article  CAS  Google Scholar 

  • Stirbet A, Govindjee (2012) Chlorophyll a fluorescence induction: a personal perspective of the thermal phase, the J-I-P rise. Photosynth Res 113:15–61

    Article  CAS  Google Scholar 

  • Strasser RJ, Govindjee (1991) The F0 and the O-J-I-P fluorescence rise in higher plants and algae. In: Argyroudi-Akoyunoglou JH (ed) Regulation of chloroplast biogenesis. Plenum Press, New York, pp 423–426

    Google Scholar 

  • Valkunas L, Liuolia V, Freiberg A (1991) Picosecond processes in chromatophores at various excitation intensities. Photosynth Res 27:83–95

    Article  CAS  Google Scholar 

  • van Best JA (1977) Studies on primary reactions of system II of photosynthesis by means of luminescence and fluorescence. Dissertation, Leiden University

  • van Best JA, Duysens LNM (1975) Reactions between primary and secondary acceptors of photosystem II in Chlorella pyrenoidosa under anaerobic conditions as studied by chlorophyll a fluorescence. Biochim Biophys Acta 408:154–163

    Article  Google Scholar 

  • van Gorkom HJ (1985) Electron transfer in photosystem II. Photosynth Res 6:97–112

    Article  Google Scholar 

  • van Gorkom HJ (1986) Fluorescence measurements in the study of photosystem II electron transport. In: Govindjee, Amesz J, Fork DC (eds) Light emission by plants and bacteria. Academic Press, New York, pp 267–289

    Chapter  Google Scholar 

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Acknowledgements

We would like to acknowledge technical assistance by Thomas Simon, Frank Reichel, and Ulrich Schliwa during the development of the MULTI-COLOR-PAM measuring system. Tony Larkum is thanked for reading the manuscript and fruitful discussions. Two anonymous reviewers are thanked for expert comments and helpful suggestions.

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Authors and Affiliations

  1. Julius-von-Sachs Institut für Biowissenschaften, Universität Würzburg, Julius-von-Sachs Platz 2, 97082, Würzburg, Germany

    Ulrich Schreiber & Christof Klughammer

  2. Heinz Walz GmbH, Eichenring 6, 91090, Effeltrich, Germany

    Gert Schansker

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  1. Ulrich Schreiber
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  2. Christof Klughammer
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  3. Gert Schansker
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Correspondence to Ulrich Schreiber.

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Schreiber, U., Klughammer, C. & Schansker, G. Rapidly reversible chlorophyll fluorescence quenching induced by pulses of supersaturating light in vivo. Photosynth Res 142, 35–50 (2019). https://doi.org/10.1007/s11120-019-00644-7

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