Abstract
In response to changes in the reduction state of the plastoquinone pool in its thylakoid membrane, the green alga Chlamydomonas reinhardtti is performing state transitions: remodelling of its thylakoid membrane leads to a redistribution of excitations over photosystems I and II (PSI and PSII). These transitions are accompanied by marked changes in the 77 K fluorescence spectrum, which form the accepted signature of state transitions. The changes are generally thought to reflect a redistribution of light-harvesting complexes (LHCs) over PSII (fluorescing below 700 nm) and PSI (fluorescing above 700 nm). Here we studied the picosecond fluorescence properties of C. reinhardtti over a broad range of wavelengths with very low excitation intensities (0.2 nJ per laser pulse). Cells were directly used for time-resolved fluorescence measurements at 77 K without further treatment, such as medium exchange with glycerol. It is observed that upon going from state 1 (relatively more fluorescence below 700 nm) to state 2 (relatively more fluorescence above 700 nm), a large part of the fluorescence of LHC/PSII becomes substantially quenched in concurrence with LHC detachment from PSII, whereas the absolute amount of PSI fluorescence hardly changes. These results are in agreement with the recent proposal that the amount of LHC moving from PSII to PSI upon going from state 1 to state 2 is rather limited (Unlu et al. Proc Natl Acad Sci USA 111 (9):3460–3465, 2014).
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References
Allen JF (1992) Protein-phosphorylation in regulation of photosynthesis. Biochim Biophys Acta 1098:275–335
Barzda V, de Grauw CJ, Vroom J, Kleima FJ, van Grondelle R, van Amerongen H, Gerritsen HC (2001) Fluorescence lifetime heterogeneity in aggregates of LHCII revealed by time-resolved microscopy. Biophys J 81:538–546
Bellafiore S, Barneche F, Peltier G, Rochaix JD (2005) State transitions and light adaptation require chloroplast thylakoid protein kinase STN7. Nature 433:892–895
Busch A, Hippler M (2011) The structure and function of eukaryotic photosystem I. Biochimica Et Biophysica Acta Bioenerg 1807:864–877
Cho F, Govindjee (1970a) Fluorescence spectra of Chlorella in the 295–77 degree K range. Biochim Biophys Acta 205:371–378
Cho F, Govindjee (1970b) Low-temperature (4–77 degrees K) spectroscopy of Anacystis: temperature dependence of energy transfer efficiency. Biochim Biophys Acta 216:151–161
Croce R, van Amerongen H (2013) Light-harvesting in photosystem I. Photosynth Res 116:153–166
Croce R, van Amerongen H (2014) Natural strategies for photosynthetic light harvesting. Nat Chem Biol 10:492–501
Delosme R, Olive J, Wollman FA (1996) Changes in light energy distribution upon state transitions: an in vivo photoacoustic study of the wild type and photosynthesis mutants from Chlamydomonas reinhardtii. Biochimica Et Biophysica Acta Bioenerg 1273:150–158
Depege N, Bellafiore S, Rochaix JD (2003) Role of chloroplast protein kinase Stt7 in LHCII phosphorylation and state transition in Chlamydomonas. Science 299:1572–1575
Drop B, Yadav KNS, Boekema EJ, Croce R (2014) Consequences of state transitions on the structural and functional organization of photosystem I in the green alga Chlamydomonas reinhardtii. Plant J 78:181–191
Finazzi G, Barbagallo RP, Bergo E, Barbato R, Forti G (2001) Photoinhibition of Chlamydomonas reinhardtii in state 1 and state 2—damages to the photosynthetic apparatus under linear and cyclic electron flow. J Biol Chem 276:22251–22257
Fleischmann MM, Ravanel S, Delosme R, Olive J, Zito F, Wollman FA, Rochaix JD (1999) Isolation and characterization of photoautotrophic mutants of Chlamydomonas reinhardtii deficient in state transition. J Biol Chem 274:30987–30994
Forti G, Caldiroli G (2005) State transitions in Chlamydomonas reinhardtii. The role of the Mehler reaction in state 2-to-state 1 transition. Plant Physiol 137:492–499
Gobets B, van Grondelle R (2001) Energy transfer and trap** in photosystem I. Biochimica Et Biophysica Acta Bioenerg 1507:80–99
Gorman DS, Levine RP (1965) Cytochrome F and Plastocyanin—their sequence in photosynthetic electron transport chain of Chlamydomonas Reinhardtti. Proc Natl Acad Sci USA 54:1665–9000
Iwai M, Minagawa J (2007) Dissociation of light-harvesting complex II from photosystem II supercomplex during state transitions in Chlamydomonas reinhardtii. Photosynth Res 91:252
Iwai M, Takahashi Y, Minagawa J (2008) Molecular remodeling of photosystem II during state transitions in Chlamydomonas reinhardtii. Plant Cell 20:2177–2189
Iwai M, Yokono M, Inada N, Minagawa J (2010) Live-cell imaging of photosystem II antenna dissociation during state transitions. Proc Natl Acad Sci USA 107:2337–2342
Kargul J, Turkina MV, Nield J, Benson S, Vener AV, Barber J (2005) Light-harvesting complex II protein CP29 binds to photosystem I of Chlamydomonas reinhardtii under state 2 conditions. FEBS J 272:4797–4806
Laptenok SP, Borst JW, Mullen KM, van Stokkum IHM, Visser AJWG, van Amerongen H (2010) Global analysis of forster resonance energy transfer in live cells measured by fluorescence lifetime imaging microscopy exploiting the rise time of acceptor fluorescence. Phys Chem Chem Phys 12:7593–7602
Lin S, Knox RS (1991) Studies of excitation energy transfer within the green alga Chlamydomonas reinhardtii and its mutants at 77 K. Photosynth Res 27:157–168
Melkozernov AN, Lin S, Blankenship RE (2000) Excitation dynamics and heterogeneity of energy equilibration in the core antenna of photosystem I from the Cyanobacterium synechocystis sp. PCC 6803. Biochemistry 39:1489–1498
Minagawa J (2011) State transitions—The molecular remodeling of photosynthetic supercomplexes that controls energy flow in the chloroplast. Biochimica Et Biophysica Acta Bioenerg 1807:897–905
Mullen KM, van Stokkum IHM (2007) TIMP: An R package for modeling multi-way spectroscopic measurements. J Stat Softw 18(3):1–46
Murata N (1969) Control of excitation transfer in photosynthesis I. Light-induced change of chlorophyll a fluoresence in Porphyridium cruentum. Biochimica et Biophysica Acta (BBA) Bioenerg 172:242–251
Nagy G, Unnep R, Zsiros O, Tokutsu R, Takizawa K, Porcar L, Moyet L, Petroutsos D, Garab G, Finazzi G, Minagawa J (2014) Chloroplast remodeling during state transitions in Chlamydomonas reinhardtii as revealed by noninvasive techniques in vivo. Proc Natl Acad Sci USA 111:5042–5047
Nelson N, Yocum CF (2006) Structure and function of photosystems I and II. Annu Rev Plant Biol 57:521–565
Palacios MA, Standfuss J, Vengris M, van Oort BF, van Stokkum IHM, Kuhlbrandt W, van Amerongen H, van Grondelle R (2006) A comparison of the three isoforms of the light-harvesting complex II using transient absorption and time-resolved fluorescence measurements. Photosynth Res 88:269–285
Schatz GH, Brock H, Holzwarth AR (1988) Kinetic and energetic model for the primary processes in photosystem-II. Biophys J 54:397–405
Snellenburg JJ, Laptenok SP, Seger R, Mullen KM, van Stokkum IHM (2012) Glotaran: a java-based graphical user interface for the R package TIMP. J Stat Softw 49:1–22
Takahashi H, Iwai M, Takahashi Y, Minagawa J (2006a) Identification of the mobile light-harvesting complex II polypeptides for state transitions. Plant Cell Physiol 47:S105
Takahashi H, Iwai M, Takahashi Y, Minagawa J (2006b) Identification of the mobile light-harvesting complex II polypeptides for state transitions in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 103:477–482
Takahashi H, Clowez S, Wollman FA, Vallon O, Rappaport F (2013) Cyclic electron flow is redox-controlled but independent of state transition. Nat Commun 4:1–8, Article ID 1954. doi:10.1038/ncomms2954
Unlu C, Drop B, Croce R, van Amerongen H (2014) State transitions in Chlamydomonas reinhardtii strongly modulate the functional size of photosystem II but not of photosystem I. Proc Natl Acad Sci USA 111:3460–3465
van Amerongen H, Croce R (2013) Light harvesting in photosystem II. Photosynth Res 116:251–263
Van Amerongen H, Valkunas L, Van Grondelle R (2000) Photosynthetic excitons. World Scientific, Singapore
van Oort B, van Hoek A, Ruban AV, van Amerongen H (2007) Aggregation of light-harvesting complex II leads to formation of efficient excitation energy traps in monomeric and trimeric complexes. FEBS Lett 581:3528–3532
van Oort B, Amunts A, Borst JW, van Hoek A, Nelson N, van Amerongen H, Croce R (2008) Picosecond fluorescence of intact and dissolved PSI–LHCI crystals. Biophys J 95:5851–5861
Van Oort B, Murali S, Wientjes E, Koehorst RBM, Spruijt RB, van Hoek A, Croce R, van Amerongen H (2009) Ultrafast resonance energy transfer from a site-specifically attached fluorescent chromophore reveals the folding of the N-terminal domain of CP29. Chem Phys 357:113–119
van Stokkum IHM, Larsen DS, van Grondelle R (2004) Global and target analysis of time-resolved spectra. Biochimica Et Biophysica Acta Bioenerg 1658:262
Van Stokkum IH, Van Oort B, Van Mourik F, Gobets B, Van Amerongen H (2008) (Sub)-Picosecond spectral evolution of fluorescence studied with a synchroscan streak-camera system and target analysis Biophysical techniques in photosynthesis. Springer, Netherlands, pp 223–240
Wientjes E, van Stokkum IH, van Amerongen H, Croce R (2011) The role of the individual Lhcas in photosystem I excitation energy trap**. Biophys J 101:745–754
Wientjes E, van Amerongen H, Croce R (2013) LHCII is an antenna of both photosystems after long-term acclimation. Biochim Biophys Acta 1827:420–426
Williams WP, Allen JF (1987) State-1/state-2 changes in higher-plants and algae. Photosynth Res 13:19–45
Wlodarczyk LM, Snellenburg JJ, Ihalainen JA, van Grondelle R, van Stokkum IH, Dekker JP (2015) Functional rearrangement of the light-harvesting antenna upon state transitions in a green alga. Biophys J 108:261–271
Wollman FA, Delepelaire P (1984) Correlation between changes in light energy-distribution and changes in thylakoid membrane polypeptide phosphorylation in Chlamydomonas reinhardtii. J Cell Biol 98:1–7
Acknowledgments
This work was supported financially by The Netherlands Organization for Scientific Research (NWO) via the Council for Chemical Sciences (HvA) and also partly supported by the BioSolar Cells Programme of The Netherlands Ministry of Economic Affairs Agriculture and Innovation and by the Foundation for Fundamental Research on Matter (FOM).
The authors would like to thank Roberta Croce (VU University Amsterdam) for useful discussions and financial support for I.P.
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Ünlü, C., Polukhina, I. & van Amerongen, H. Origin of pronounced differences in 77 K fluorescence of the green alga Chlamydomonas reinhardtii in state 1 and 2. Eur Biophys J 45, 209–217 (2016). https://doi.org/10.1007/s00249-015-1087-9
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DOI: https://doi.org/10.1007/s00249-015-1087-9