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.
Similar content being viewed by others
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
Breton J, Geacintov NA, Swenberg CE (1979) Quenching of fluorescence by triplet excited states in chloroplasts. Biochim Biophys Acta 548:616–635
Butler WL (1972) On the primary nature of fluorescence yield changes associated with photosynthesis. Proc Natl Acad Sci USA 69:3420–3422
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
Papageorgiou GC, Govindjee (eds) (2004) Chlorophyll fluorescence: a signature of photosynthesis. Springer, Dordrecht
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
Pfündel EE (1998) Estimating the contribution of photosystem I to total leaf chlorophyll fluorescence. Photosynth Res 56:185–195
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
Pirson A, Ruppel HG (1962) Über die Induktion einer Teilungshemmung in synchronen Kulturen von Chlorella. Arch Mikrobiol 42:499–505
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
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
Schansker G, Tóth SZ, Holzwarth AR (2014) Chlorophyll a fluorescence: beyond the limits of the QA model. Photosynth Res 120:43–58
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
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
Schreiber U, Vidaver W (1974) Chlorophyll fluorescence induction in anaerobic Scenedesmus obliquus. Biochim Biophys Acta 368:97–112
Schreiber U, Vidaver W (1975) Analysis of anaerobic fluorescence decay in Scenedesmus obliquus. Biochim Biophys Acta 387:37–51
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
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
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
Seaton GGR, Walker DA (1990) Chlorophyll fluorescence as a measure of photosynthetic carbon assimilation. Proc R Soc Lond 242:29–35
Siefermann-Harms (1987) The light-harvesting and protective functions of carotenoids in photosynthetic membranes. Physiol Plant 69:561–568
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
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
Valkunas L, Liuolia V, Freiberg A (1991) Picosecond processes in chromatophores at various excitation intensities. Photosynth Res 27:83–95
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
van Gorkom HJ (1985) Electron transfer in photosystem II. Photosynth Res 6:97–112
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
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.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11120-019-00644-7