Abstract
PAM fluorescence of leaves of cherry laurel (Prunus laurocerasus L.) was measured simultaneously in the spectral range below 700 nm (sw) and above 700 nm (lw). A high-sensitivity photodiode was employed to measure the low intensities of sw fluorescence. Photosystem II (PSII) performance was analyzed by the saturation pulse method during a light response curve with subsequent dark phase. The sw fluorescence was more variable, resulting in higher PSII photochemical yields compared to lw fluorescence. The variations between sw and lw data were explained by different levels of photosystem I (PSI) fluorescence: the contribution of PSI fluorescence to minimum fluorescence (F0) was calculated to be 14% at sw wavelengths and 45% at lw wavelengths. With the results obtained, the validity of an earlier method for the quantification of PSI fluorescence (Genty et al. in Photosynth Res 26:133–139, 1990, https://doi.org/10.1007/BF00047085) was reconsidered. After subtracting PSI fluorescence from all fluorescence levels, the maximum PSII photochemical yield (FV/FM) in the sw range was 0.862 and it was 0.883 in the lw range. The lower FV/FM at sw wavelengths was suggested to arise from inactive PSII reaction centers in the outermost leaf layers. Polyphasic fluorescence transients (OJIP or OI1I2P kinetics) were recorded simultaneously at sw and lw wavelengths: the slowest phase of the kinetics (IP or I2P) corresponded to 11% and 13% of total variable sw and lw fluorescence, respectively. The idea that this difference is due to variable PSI fluorescence is critically discussed. Potential future applications of simultaneously recording fluorescence in two spectral windows include studies of PSI non-photochemical quenching and state I–state II transitions, as well as measuring the fluorescence from pH-sensitive dyes simultaneously with chlorophyll fluorescence.
This is a preview of subscription content, log in via an institution to check access.
Availability of data and materials
Original data will be provided on request.
Abbreviations
- sw:
-
Short wavelength range (< 700 nm)
- lw:
-
Long wavelength range (> 700 nm)
References
Baker NR (2008) Chlorophyll fluorescence: a probe of photosynthesis in vivo. Annu Rev Plant Biol 59:89–113. https://doi.org/10.1146/annurev.arplant.59.032607.092759
Baker NR, Oxborough K (2004) Chlorophyll fluorescence as a probe of photosynthetic productivity. In: Papageorgiou GC, Govindjee (eds) Chlorophyll a fluorescence: a signature of photosynthesis, Springer, Dordrecht, p 65–82. https://doi.org/10.1007/978-1-4020-3218-9_3
Ballottari M, Alcocer MJP, D’Andrea C, Viola D, Ahn TK, Petrozza A, Polli D, Fleming GR, Cerullo G, Bassi R (2014) Regulation of photosystem I light harvesting by zeaxanthin. Proc Natl Acad Sci USA 111:E2431–E2438. https://doi.org/10.1073/pnas.1404377111
Bilger W, Björkman O (1990) Role of the xanthophyll cycle in photoprotection elucidated by measurements of light-induced absorbance changes, fluorescence and photosynthesis in leaves of Hedera canariensis. Photosynth Res 25:173–185. https://doi.org/10.1007/BF00033159
Brugnoli E, Björkman O (1992) Chloroplast movements in leaves: influence on chlorophyll fluorescence and measurements of light-induced absorbance changes related to ΔpH and zeaxanthin formation. Photosynth Res 32:23–35. https://doi.org/10.1007/BF00028795
Buschmann C, Lenk S, Lichtenthaler HK (2012) Reflectance spectra and images of green leaves with different tissue structure and chlorophyll content. Isr J Plant Sci 60:49–64. https://doi.org/10.1560/IJPS.60.1-2.49
Byrdin M, Rimke I, Schlodder E, Stehlik D, Roelofs TA (2000) Decay kinetics and quantum yields of fluorescence in Photosystem I from Synechococcus elongatus with P700 in the reduced and oxidized state: are the kinetics of excited state decay trap-limited or transfer-limited? Biophys J 79:992–1007. https://doi.org/10.1016/S0006-3495(00)76353-3
Cho F, Govindjee (1970) Low-temperature (4–77°K) spectroscopy of chlorella; temperature dependence of energy transfer efficiency. Biochim Biophys Acta 216:139–150. https://doi.org/10.1016/0005-2728(70)90166-0
Chukhutsina VU, Holzwarth AR, Croce R (2019) Time-resolved fluorescence measurements on leaves: principles and recent developments. Photosynth Res 140:355–369. https://doi.org/10.1007/s11120-018-0607-8
Chylla RA, Whitmarsh J (1989) Inactive photosystem II complexes in leaves Turnover rate and quantitation. Plant Physiol 90:765–772. https://doi.org/10.1104/pp.90.2.765
Dall’Osto L, Cazzaniga S, Wada M, Bassi R (2014) On the origin of a slowly reversible fluorescence decay component in the Arabidopsis npq4 mutant. Phil Trans R Soc Lond B 369:20130221. https://doi.org/10.1098/rstb.2013.0221
Dekker JP, Boekema EJ (2005) Supramolecular organization of thylakoid membrane proteins in green plants. Biochim Biophys Acta 1706:12–39. https://doi.org/10.1016/j.bbabio.2004.09.009
Duysens LNM (1951) Transfer of light energy within the pigment systems present in photosynthesizing cells. Nature 168:548–550. https://doi.org/10.1038/168548a0
Esteban R, Barrutia O, Artetxe U, Fernández-Marín B, Hernández A, García-Plazaola JI (2015) Internal and external factors affecting photosynthetic pigment composition in plants: a meta-analytical approach. New Phytol 206:268–280. https://doi.org/10.1111/nph.13186
Forster LS, Livingston R (1952) The absolute quantum yields of the fluorescence of chlorophyll solutions. J Chem Phys 20:1315–1320. https://doi.org/10.1063/1.1700727
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. https://doi.org/10.1016/S0005-2728(02)00366-3
Gates DM, Keegan HJ, Schleter JC, Weidner VR (1965) Spectral properties of plants. Appl Opt 4:11–20. https://doi.org/10.1364/AO.4.000011
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. https://doi.org/10.1016/S0304-4165(89)80016-9
Genty B, Wonders J, Baker NR (1990) Non-photochemical quenching of F0 in leaves is emission wavelength dependent. Consequences for quenching analysis and its interpretation. Photosynth Res 26:133–139. https://doi.org/10.1007/BF00047085
Gilmore AM, Yamamoto HY (1991) Zeaxanthin formation and energy-dependent fluorescence quenching in pea chloroplasts under artificially mediated linear and cyclic electron transport. Plant Physiol 96:635–643. https://doi.org/10.1104/pp.96.2.635
Giovagnetti V, Ware MA, Ruban AV (2015) Assessment of the impact of photosystem I chlorophyll fluorescence on the pulse-amplitude modulated quenching analysis in leaves of Arabidopsis thaliana. Photosynth Res 125:179–189. https://doi.org/10.1007/s11120-015-0087-z
Girolomoni L, Bellamoli F, de la Cruz VG, Perozeni F, D’Andrea C, Cerullo G, Cazzaniga S, Ballottari M (2020) Evolutionary divergence of photoprotection in the green algal lineage: a plant-like violaxanthin de-epoxidase enzyme activates the xanthophyll cycle in the green alga Chlorella vulgaris modulating photoprotection. New Phytol. https://doi.org/10.1111/nph.16674
Haldrup A, Jensen PE, Lunde C, Scheller HV (2001) Balance of power: a view of the mechanism of photosynthetic state transitions. Trends Plant Sci 6:301–305. https://doi.org/10.1016/S1360-1385(01)01953-7
Johnson MP, Ruban AV (2009) Photoprotective energy dissipation in higher plants involves alteration of the excited state energy of the emitting chlorophyll(s) in the light harvesting antenna II (LHCII). J Biol Chem 284:23592–23601. https://doi.org/10.1074/jbc.m109.013557
Kasahara M, Kagava T, Oikava K, Suetsugu N, Miyao M, Wada M (2002) Chloroplast avoidance movement reduces photodamage in plants. Nature 420:829–832. https://doi.org/10.1038/nature01213
Keren N, Berg A, van Kan PJM, Levanon H, Ohad I (1997) Mechanism of photosystem II photoinactivation and D1 protein degradation at low light: the role of back electron flow. Proc Natl Acad Sci USA 94:1579–1584. https://doi.org/10.1073/pnas.94.4.1579
Kitajima M, Butler WL (1975) Quenching of chlorophyll fluorescence and primary photochemistry in chloroplasts by dibromothymoquinone. Biochim Biophys Acta 376:105–115. https://doi.org/10.1016/0005-2728(75)90209-1
Krause GH, Jahns P (2004) Non-photochemical energy dissipation determined by chlorophyll fluorescence quenching: characterization and function. In: Papageorgiou GC, Govindjee (eds) Chlorophyll a fluorescence. A signature of photosynthesis. Springer, Berlin, p 463–495. https://doi.org/10.1007/978-1-4020-3218-9_18
Kume A (2017) Importance of the green color, absorption gradient, and spectral absorption of chloroplasts for the radiative energy balance of leaves. J Plant Res 130:501–514. https://doi.org/10.1007/s10265-017-0910-Z
Laisk A, Oja V (2020) Variable fluorescence of closed photochemical reaction centers. Photosynth Res 143:335–346. https://doi.org/10.1007/s11120-020-00712-3
Lavergne J, Leci E (1993) Properties of inactive Photosystem II centers. Photosynth Res 35:323–343. https://doi.org/10.1007/BF00016563
Lazár D (2013) Simulations show that a small part of variable chlorophyll a fluorescence originates in photosystem I and contributes to overall fluorescence rise. J Theor Biol 335:249–264. https://doi.org/10.1016/j.jtbi.2013.06.028
Lazár D, Schansker G (2009) Models of chlorophyll a fluorescence transients. In: Laisk A, Nedbal L, Govindjee (eds) Understanding complexity from molecules to ecosystems. Springer, Dordrecht, p 85–123. https://doi.org/10.1007/978-1-4020-9237-4_5
Magyar M, Sipka G, Kovács L, Ughy B, Zhu Q, Han G, Špunda V, Lambrev PH, Shen J-R, Garab G (2018) Rate-limiting steps in the dark-to-light transition of Photosystem II—revealed by chlorophyll-a fluorescence induction. Sci Rep 8:2755. https://doi.org/10.1038/s41598-018-21195-2
Mamedov M, Govindjee NV, Semenov A (2015) Primary electron transfer processes in photosynthetic reaction centers from oxygenic organisms. Photosynth Res 125:51–63. https://doi.org/10.1007/s11120-015-0088-y
Nilkens M, Kress E, Lambrev P, Miloslavina Y, Müller M, Holzwarth AR, Jahns P (2010) Identification of a slowly inducible zeaxanthin-dependent component of non-photochemical quenching of chlorophyll fluorescence generated under steady-state conditions in Arabidopsis. Biochim Biophys Acta 1797:466–475. https://doi.org/10.1016/j.bbabio.2010.01.001
Oxborough K, Baker NR (1997) Resolving chlorophyll a fluorescence images of photosynthetic efficiency into photochemical and nonphotochemical components—calculation of qP and F′v/F′m without measuring F′o. Photosynth Res 54:135–142. https://doi.org/10.1023/A:1005936823310
Park Y-I, Chow WS, Anderson JM (1995) The quantum yield of photoinactivation of photosystem II in pea leaves is greater at low than high photon exposure. Plant Cell Physiol 36:1163–1167. https://doi.org/10.1093/oxfordjournals.pcp.a078863
Peterson RB, Oja V, Laisk A (2001) Chlorophyll fluorescence at 680 and 730 nm and leaf photosynthesis. Photosynth Res 70:185–196. https://doi.org/10.1023/A:1017952500015
Peterson RB, Oja V, Eichelmann H, Bichele I, Dall’Osto L, Laisk A (2014) Fluorescence F0 of photosystems II and I in developing C3 and C4 leaves, and implications on regulation of excitation balance. Photosynth Res 122:41–56. https://doi.org/10.1007/s11120-014-0009-5
Pfündel EE (1998) Estimating the contribution of photosystem I to total leaf chlorophyll fluorescence. Photosynth Res 56:185–195. https://doi.org/10.1023/A:1006032804606
Pfündel EE, Agati G, Cerovic ZG (2008) Optical properties of plant surfaces. In: Riederer M, Müller C (eds) Biology of the plant cuticle. Annual plant reviews, vol 23. Blackwell Publishing, Oxford, p 216–249. https://doi.org/10.1002/9781119312994.apr0234
Pfündel EE, Klughammer C, Meister A, Cerovic ZG (2013) Deriving fluorometer-specific values of relative PSI fluorescence intensity from quenching of F0 fluorescence in leaves of Arabidopsis thaliana and Zea mays. Photosynth Res 114:189–206. https://doi.org/10.1007/s11120-012-9788-8
Pfündel EE, Latouche G, Meister A, Cerovic ZG (2018) Linking chloroplast relocation to different responses of photosynthesis to blue and red radiation in low and high light-acclimated leaves of Arabidopsis thaliana (L.). Photosynth Res 137:105–128. https://doi.org/10.1007/s11120-018-0482-3
Pospíšil P, Dau H (2002) Valinomycin sensitivity proves that light-induced thylakoid voltages result in millisecond phase of chlorophyll fluorescence transients. Biochim Biophys Acta 1554:94–100. https://doi.org/10.1016/S0005-2728(02)00216-5
Quick WP, Stitt M (1989) An examination of factors contribution to non-photochemical quenching of chlorophyll fluorescence in barley leaves. Biochim Biophys Acta 977:287–296. https://doi.org/10.1016/S0005-2728(89)80082-9
Remelli W, Santabarbara S (2018) Excitation and emission wavelength dependence of fluorescence spectra in whole cells of the cyanobacterium Synechocystis sp. PPC6803: influence on the estimation of Photosystem II maximal quantum efficiency. Biochim Biophys Acta 1859:1207–1222. https://doi.org/10.1016/j.bbabio.2018.09.366
Roelofs TA, Lee CH, Holzwarth AR (1992) Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. A new approach to the characterization of the primary processes in photosystem II α- and β-units. Biophys J 61:1147–1163. https://doi.org/10.1016/S0006-3495(92)81924-0
Ruban AV, Johnson MP (2015) Visualizing the dynamic structure of the plant photosynthetic membrane. Nat Plants 1:15161. https://doi.org/10.1038/nplants.2015.161
Santabarbara S, Monteleone FV, Remelli W, Rizzo F, Menin B, Casazza AP (2019) Comparative excitation-emission dependence of the FV/FM ratio in model green algae and cyanobacterial strains. Physiol Plant 166:351–364. https://doi.org/10.1111/ppl.12931
Schansker G, Strasser RJ (2005) Quantification of non-QB-reducing centers in leaves using a far-red pre-illumination. Photosynth Res 84:145–151. https://doi.org/10.1007/s11120-004-7156-z
Schansker G, Tóth SZ, Kovács L, Holzwarth AR, Garab G (2011) Evidence for a fluorescence yield change driven by a light-induced conformational change within photosystem II during the fast chlorophyll a fluorescence rise. Biochim Biophys Acta 1807:1032–1043. https://doi.org/10.1016/j.bbabio.2011.05.022
Schlodder E, Çetin M, Byrdin M, Terekhova I, Karapetyan NV (2005) P700+- and 3P700-induced quenching of the fluorescence at 760 nm in trimeric Photosystem I complexes from the cyanobacterium Arthrospira platensis. Biochim Biophys Acta 1706:53–67. https://doi.org/10.1016/j.bbabio.2004.08.009
Schreiber U (2004) Pulse-amplitude-modulation (PAM) fluorometry and saturation pulse method: an overview. In: Papageorgiou GC, Govindjee (eds) Chlorophyll a fluorescence: a signature of photosynthesis. Springer, Berlin, p 279–319. https://doi.org/10.1007/978-1-4020-3218-9_11
Schreiber U, Klughammer C (2021) Evidence for variable chlorophyll fluorescence of photosystem I in vivo. Photosynth Res. https://doi.org/10.1007/s11120-020-00814-y
Schreiber U, Schliwa U, Bilger W (1986) Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulation fluorometer. Photosynth Res 10:51–62. https://doi.org/10.1007/BF00024185
Schreiber U, Neubauer C, Klughammer C (1989) Devices and methods for room-temperature fluorescence analysis. Philos Trans R Soc Lond B 323:241–251. https://doi.org/10.1098/rstb.1989.0007
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. https://doi.org/10.1071/PP9950209
Stirbet A, Govindjee (2011) On the relation between the Kautsky effect (chlorophyll a fluorescence induction) and Photosystem II: basics and applications of the OJIP fluorescence transient. J Photochem Photobiol B 104:236–257. https://doi.org/10.1016/j.jphotobiol.2010.12.010
Stirbet A, Govindjee (2012) Chlorophyll a fluorescence induction: a personal perspective of the thermal phase, the JIP rise. Photosynth Res 113:15–61. https://doi.org/10.1007/s11120-012-9754-5
Strasser RJ, Srivastava A, Govindjee (1995) Polyphasic chlorophyll a fluorescence transients in plants and cyanobacteria. Photochem Photobiol 61:32–42. https://doi.org/10.1111/j.1751-1097.1995.tb09240.x
Tian L, Xu P, Chukhutsina VU, Holzwarth AR, Croce R (2017) Zeaxanthin-dependent nonphotochemical quenching does not occur in photosystem I in the higher plant Arabidopsis thaliana. Proc Natl Acad Sci USA 114:4828–4832. https://doi.org/10.1073/pnas.1621051114
Trissl HW (1997) Determination of the quenching efficiency of the oxidized primary donor of Photosystem I, P700+: implications for the trapping mechanism. Photosynth Res 54:237–240. https://doi.org/10.1023/A:1005981016835
Trissl H-W, Gao Y, Wulf K (1993) Theoretical fluorescence induction curves derived from coupled differential equations describing the primary photochemistry of photosystem II by an exciton-radical pair equilibrium. Biophys J 64:974–988. https://doi.org/10.1016/S0006-3495(93)81463-2
van Grondelle R, Dekker JP, Gillbro T, Sundstrom V (1994) Energy transfer and trapping in photosynthesis. Biochim Biophys Acta 1187:1–65. https://doi.org/10.1016/0005-2728(94)90166-X
Van Kooten O, Snel JFH (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth Res 2:147–150. https://doi.org/10.1007/BF00033156
Weber G, Teale FWJ (1957) Determination of the absolute quantum yield of fluorescent solutions. Trans Faraday Soc 53:646–655. https://doi.org/10.1039/TF9575300646
Weis E (1985) Chlorophyll fluorescence at 77 K in intact leaves: characterization of a technique to eliminate artifacts related to self-absorption. Photosynth Res 6:73–86. https://doi.org/10.1007/BF00029047
Wientjes E, Croce R (2012) PMS: photosystem I electron donor or fluorescence quencher. Photosynth Res 111:185–191. https://doi.org/10.1007/s11120-011-9671-z
Wientjes E, van Amerongen H, Croce R (2013) Quantum yield of charge separation in photosystem II: functional effect of changes in the antenna size upon light acclimation. J Phys Chem 117:11200–11208. https://doi.org/10.1021/jp401663w
Yin ZH, Neimanis S, Heber U (1990a) Light-dependent pH changes in leaves of C3 plants. 2. Effect of CO2 and O2 on the cytosolic and the vacuolar pH. Planta 182:253–261. https://doi.org/10.1007/bf00197119
Yin ZH, Neimanis S, Wagner U, Heber U (1990b) Light-dependent pH changes in leaves of C3 plants. 1. Recording pH changes in various cellular compartments by fluorescent probes. Planta 182:244–252. https://doi.org/10.1007/bf00197118
Zhang Y, Huang J, Wang F, Blackburn GA, Zhang HK, Wang X, Wei C, Zhang K, Wei C (2017) An extended PROSPECT: advance in the leaf optical properties model separating total chlorophylls into chlorophyll a and b. Sci Rep 7:6429. https://doi.org/10.1038/s41598-017-06694-y
Acknowledgements
The outstanding technical support by Frank Reichel and Thomas Simon is greatly acknowledged. I thank Todd Kana for proofreading. Specials thanks go to Gert Schansker for fruitful discussions and essential support during the preparation of this paper. I am particularly grateful to Christof Klughammer for implementing the new fluorescence option in the DualPAM software.
Funding
Does not apply.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The author states that there is no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Pfündel, E.E. Simultaneously measuring pulse-amplitude-modulated (PAM) chlorophyll fluorescence of leaves at wavelengths shorter and longer than 700 nm. Photosynth Res 147, 345–358 (2021). https://doi.org/10.1007/s11120-021-00821-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11120-021-00821-7