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Dynamic thylakoid stacking and state transitions work synergistically to avoid acceptor-side limitation of photosystem I

A Publisher Correction to this article was published on 29 January 2021

This article has been updated

Abstract

TAP38/STN7-dependent (de)phosphorylation of light-harvesting complex II (LHCII) regulates the relative excitation rates of photosystems I and II (PSI, PSII) (state transitions) and the size of the thylakoid grana stacks (dynamic thylakoid stacking). Yet, it remains unclear how changing grana size benefits photosynthesis and whether these two regulatory mechanisms function independently. Here, by comparing Arabidopsis wild-type, stn7 and tap38 plants with the psal mutant, which undergoes dynamic thylakoid stacking but lacks state transitions, we explain their distinct roles. Under low light, smaller grana increase the rate of PSI reduction and photosynthesis by reducing the diffusion distance for plastoquinol; however, this beneficial effect is only apparent when PSI/PSII excitation balance is maintained by state transitions or far-red light. Under high light, the larger grana slow plastoquinol diffusion and lower the equilibrium constant between plastocyanin and PSI, maximizing photosynthesis by avoiding PSI photoinhibition. Loss of state transitions in low light or maintenance of smaller grana in high light also both bring about a decrease in cyclic electron transfer and over-reduction of the PSI acceptor side. These results demonstrate that state transitions and dynamic thylakoid stacking work synergistically to regulate photosynthesis in variable light.

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Fig. 1: Changes in thylakoid grana diameter and excitation energy distribution between photosystems in LL and HL.
Fig. 2: Photosynthetic properties of WT, tap38, stn7 and psal Arabidopsis plants determined by infrared gas exchange, chlorophyll fluorescence and P700 absorption spectroscopy.
Fig. 3: DIRK analysis of the reduction PSI (P700+), Pc+ and oxidation of Fd.
Fig. 4: Investigating possible causes of defective electron transfer regulation in stn7 and tap38.
Fig. 5: Comparison of PSI acceptor-side limitation and CET.

Data availability

The datasets analysed during the current study are available from the corresponding author on reasonable request. The sequence data from this article can be found in The Arabidopsis Information Resource or GenBank/EMBL database under the following accession numbers: STN7 (At1g68830), TAP38/PPH1 (At4t27800), CURT1A (At4g01150), CURT1B (At2g46820), CURT1C (At1g52220), CURT1D (At4g38100) and PSAL (At4g12800).

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References

  1. 1.

    Ruban, A. V. Evolution under the sun: optimizing light harvesting in photosynthesis. J. Exp. Bot. https://doi.org/10.1093/jxb/eru400 (2015).

  2. 2.

    Miyake, C. Molecular mechanism of oxidation of p700 and suppression of ROS production in photosystem I in response to electron-sink limitations in C3 plants. Antioxidants 9, 230 (2020).

    CAS  Article  Google Scholar 

  3. 3.

    Li, Z., Wakao, S., Fischer, B. B. & Niyogi, K. K. Sensing and responding to excess light. Annu. Rev. Plant Biol. https://doi.org/10.1146/annurev.arplant.58.032806.103844 (2009).

  4. 4.

    Ruban, A. V., Johnson, M. P. & Duffy, C. D. P. The photoprotective molecular switch in the photosystem II antenna. Biochim. Biophys. Acta Bioenerg. 1817, 167–181 (2012).

    CAS  Article  Google Scholar 

  5. 5.

    Suorsa, M. et al. PGR5 ensures photosynthetic control to safeguard photosystem I under fluctuating light conditions. Plant Signal. Behav. 8, e22741 (2013).

    Article  Google Scholar 

  6. 6.

    Johnson, G. N. Physiology of PSI cyclic electron transport in higher plants. Biochim. Biophys. Acta Bioenerg. 1807, 384–389 (2011).

    CAS  Article  Google Scholar 

  7. 7.

    Yamori, W. & Shikanai, T. Physiological functions of cyclic electron transport around photosystem I in sustaining photosynthesis and plant growth. Annu. Rev. Plant Biol. 67, 81–106 (2016).

    CAS  Article  Google Scholar 

  8. 8.

    Hertle, A. P. et al. PGRL1 is the elusive ferredoxin-plastoquinone reductase in photosynthetic cyclic electron flow. Mol. Cell https://doi.org/10.1016/j.molcel.2012.11.030 (2013).

  9. 9.

    Nandha, B., Finazzi, G., Joliot, P., Hald, S. & Johnson, G. N. The role of PGR5 in the redox poising of photosynthetic electron transport. Biochim. Biophys. Acta Bioenerg. 1767, 1252–1259 (2007).

    CAS  Article  Google Scholar 

  10. 10.

    Joliot, P. & Johnson, G. N. Regulation of cyclic and linear electron flow in higher plants. Proc. Natl Acad. Sci. USA 108, 13317–13322 (2011).

    CAS  Article  Google Scholar 

  11. 11.

    Allen, J. F. State transitions—a question of balance. Science 299, 1530–1532 (2003).

    CAS  Article  Google Scholar 

  12. 12.

    Ruban, A. V. & Johnson, M. P. Dynamics of higher plant photosystem cross-section associated with state transitions. Photosynth. Res. 99, 173–183 (2009).

    CAS  Article  Google Scholar 

  13. 13.

    Bellaflore, S., Barneche, F., Peltler, G. & Rochalx, J. D. State transitions and light adaptation require chloroplast thylakoid protein kinase STN7. Nature 433, 892–895 (2005).

    Article  Google Scholar 

  14. 14.

    Shapiguzov, A. et al. The PPH1 phosphatase is specifically involved in LHCII dephosphorylation and state transitions in Arabidopsis. Proc. Natl Acad. Sci. USA https://doi.org/10.1073/pnas.0913810107 (2010).

  15. 15.

    Pribil, M., Pesaresi, P., Hertle, A., Barbato, R. & Leister, D. Role of plastid protein phosphatase TAP38 in LHCII dephosphorylation and thylakoid electron flow. PLoS Biol. https://doi.org/10.1371/journal.pbio.1000288 (2010).

  16. 16.

    Vener, A. V., Van Kan, P. J. M., Rich, P. R., Ohad, I. & Andersson, B. Plastoquinol at the quinol oxidation site of reduced cytochrome bf mediates signal transduction between light and protein phosphorylation: thylakoid protein kinase deactivation by a single-turnover flash. Proc. Natl Acad. Sci. USA 94, 1585–1590 (1997).

    CAS  Article  Google Scholar 

  17. 17.

    Rintamäki, E., Martinsuo, P., Pursiheimo, S. & Aro, E. M. Cooperative regulation of light-harvesting complex II phosphorylation via the plastoquinol and ferredoxin-thioredoxin system in chloroplasts. Proc. Natl Acad. Sci. USA 97, 11644–11649 (2000).

    Article  Google Scholar 

  18. 18.

    Fernyhough, P., Foyer, C. H. & Horton, P. Increase in the level of thylakoid protein phosphorylation in maize mesophyll chloroplasts by decrease in the transthylakoid pH gradient. FEBS Lett. 176, 133–138 (1984).

    CAS  Article  Google Scholar 

  19. 19.

    Taylor, C. R., Van Ieperen, W. & Harbinson, J. Demonstration of a relationship between state transitions and photosynthetic efficiency in a higher plant. Biochem. J. https://doi.org/10.1042/BCJ20190576 (2019).

  20. 20.

    Frenkel, M. et al. Improper excess light energy dissipation in Arabidopsis results in a metabolic reprogramming. BMC Plant Biol. https://doi.org/10.1186/1471-2229-9-12 (2009).

  21. 21.

    Külheim, C., Ågren, J. & Jansson, S. Rapid regulation of light harvesting and plant fitness in the field. Science 297, 91–93 (2002).

    Article  Google Scholar 

  22. 22.

    Suorsa, M. et al. Proton gradient regulation5 is essential for proper acclimation of Arabidopsis photosystem I to naturally and artificially fluctuating light conditions. Plant Cell 24, 2934–2948 (2012).

    CAS  Article  Google Scholar 

  23. 23.

    Lunde, C., Jensen, P. E., Haldrup, A., Knoetzel, J. & Scheller, H. V. The PSI-H subunit of photosystem I is essential for state transitions in plant photosynthesis. Nature https://doi.org/10.1038/35046121 (2000).

  24. 24.

    Fernyhough, P., Foyer, C. & Horton, P. The influence of metabolic state on the level of phosphorylation of the light-harvesting chlorophyll-protein complex in chloroplasts isolated from maize mesophyll. Biochim. Biophys. Acta Bioenerg. https://doi.org/10.1016/0005-2728(83)90235-9 (1983).

  25. 25.

    Allen, J. F. Protein phosphorylation—carburettor of photosynthesis? Trends Biochem. Sci. 8, 369–373 (1983).

    CAS  Article  Google Scholar 

  26. 26.

    Cardol, P. et al. Impaired respiration discloses the physiological significance of state transitions in Chlamydomonas. Proc. Natl Acad. Sci. USA 15, 15979–15984 (2009).

    Article  Google Scholar 

  27. 27.

    Bulté, L., Gans, P., Rebéillé, F. & Wollman, F. A. ATP control on state transitions in vivo in Chlamydomonas reinhardtii. Biochim. Biophys. Acta Bioenerg. https://doi.org/10.1016/0005-2728(90)90095-L (1990).

  28. 28.

    Takahashi, H., Clowez, S., Wollman, F. A., Vallon, O. & Rappaport, F. Cyclic electron flow is redox-controlled but independent of state transition. Nat. Commun. 4, 1954 (2013).

    Article  Google Scholar 

  29. 29.

    Pesaresi, P. et al. Arabidopsis STN7 kinase provides a link between short- and long-term photosynthetic acclimation. Plant Cell 21, 2402–2423 (2009).

    CAS  Article  Google Scholar 

  30. 30.

    Kyle, D. J., Staehelin, L. A. & Arntzen, C. J. Lateral mobility of the light-harvesting complex in chloroplast membranes controls excitation energy distribution in higher plants. Arch. Biochem. Biophys. 222, 527–541 (1983).

    CAS  Article  Google Scholar 

  31. 31.

    Rozak, P. R., Seiser, R. M., Wacholtz, W. F. & Wise, R. R. Rapid, reversible alterations in spinach thylakoid appression upon changes in light intensity. Plant Cell Environ. 25, 421–429 (2002).

    Article  Google Scholar 

  32. 32.

    Wood, W. H. J., Barnett, S. F. H., Flannery, S., Hunter, C. N. & Johnson, M. P. Dynamic thylakoid stacking is regulated by LHCII phosphorylation but not its interaction with PSI. Plant Physiol. 180, 2152–2166 (2019).

    CAS  Article  Google Scholar 

  33. 33.

    Wood, W. H. J. et al. Dynamic thylakoid stacking regulates the balance between linear and cyclic photosynthetic electron transfer. Nat. Plants 4, 116–127 (2018).

    CAS  Article  Google Scholar 

  34. 34.

    Anderson, J. M., Horton, P., Kim, E. H. & Chow, W. S. Towards elucidation of dynamic structural changes of plant thylakoid architecture. Phil. Trans. R. Soc. B Bio. Sci. https://doi.org/10.1098/rstb.2012.0373 (2012).

  35. 35.

    Capretti, A. et al. Nanophotonics of higher-plant photosynthetic membranes. Light Sci. Appl. https://doi.org/10.1038/s41377-018-0116-8 (2019).

  36. 36.

    Grieco, M., Tikkanen, M., Paakkarinen, V., Kangasjärvi, S. & Aro, E. M. Steady-state phosphorylation of light-harvesting complex II proteins preserves photosystem I under fluctuating white light. Plant Physiol. 160, 1896–1910 (2012).

    CAS  Article  Google Scholar 

  37. 37.

    Tikkanen, M., Grieco, M., Kangasjärvi, S. & Aro, E. M. Thylakoid protein phosphorylation in higher plant chloroplasts optimizes electron transfer under fluctuating light. Plant Physiol. https://doi.org/10.1104/pp.109.150250 (2010).

  38. 38.

    Tikkanen, M. et al. State transitions revisited—a buffering system for dynamic low light acclimation of Arabidopsis. Plant Mol. Biol. https://doi.org/10.1007/s11103-006-9088-9 (2006).

  39. 39.

    Pribil, M., Labs, M. & Leister, D. Structure and dynamics of thylakoids in land plants. J. Exp. Bot. https://doi.org/10.1093/jxb/eru090 (2014).

  40. 40.

    Schreiber, U. & Klughammer, C. Analysis of photosystem I donor and acceptor sides with a new type of online-deconvoluting kinetic LED-array spectrophotometer. Plant Cell Physiol. https://doi.org/10.1093/pcp/pcw044 (2016).

  41. 41.

    Klughammer, C. & Schreiber, U. An improved method, using saturating light pulses, for the determination of photosystem I quantum yield via P700+-absorbance changes at 830 nm. Planta https://doi.org/10.1007/BF00194461 (1994).

  42. 42.

    Klughammer, C. & Schreiber, U. Deconvolution of ferredoxin, plastocyanin, and P700 transmittance changes in intact leaves with a new type of kinetic LED array spectrophotometer. Photosynth. Res. https://doi.org/10.1007/s11120-016-0219-0 (2016).

  43. 43.

    Sacksteder, C. A. & Kramer, D. M. Dark-interval relaxation kinetics (DIRK) of absorbance changes as a quantitative probe of steady-state electron transfer. Photosynth. Res. https://doi.org/10.1023/A:1010785912271 (2000).

  44. 44.

    Kirchhoff, H., Schöttler, M. A., Maurer, J. & Weis, E. Plastocyanin redox kinetics in spinach chloroplasts: evidence for disequilibrium in the high potential chain. Biochim. Biophys. Acta Bioenerg. 1659, 63–72 (2004).

    CAS  Article  Google Scholar 

  45. 45.

    Ott, T., Clarke, J., Birks, K. & Johnson, G. Regulation of the photosynthetic electron transport chain. Planta https://doi.org/10.1007/s004250050629 (1999).

  46. 46.

    Jahns, P., Graf, M., Munekage, Y. & Shikanai, T. Single point mutation in the rieske iron-sulfur subunit of cytochrome b6/f leads to an altered pH dependence of plastoquinol oxidation in Arabidopsis. FEBS Lett. 519, 99–102 (2002).

    CAS  Article  Google Scholar 

  47. 47.

    Correa Galvis, V. et al. H+ transport by K+ EXCHANGE ANTIPORTER3 promotes photosynthesis and growth in chloroplast ATP synthase mutants. Plant Physiol. https://doi.org/10.1104/pp.19.01561 (2020).

  48. 48.

    Klughammer, C., Siebke, K. & Schreiber, U. Continuous ECS-indicated recording of the proton-motive charge flux in leaves. Photosynth. Res. https://doi.org/10.1007/s11120-013-9884-4 (2013).

  49. 49.

    Sacksteder, C. A., Kanazawa, A., Jacoby, M. E. & Kramer, D. M. The proton to electron stoichiometry of steady-state photosynthesis in living plants: a proton-pumping Q cycle is continuously engaged. Proc. Natl Acad. Sci. USA 97, 14283–14288 (2000).

    CAS  Article  Google Scholar 

  50. 50.

    Johnson, M. P. & Ruban, A. V. Rethinking the existence of a steady-state Δψ component of the proton motive force across plant thylakoid membranes. Photosynth. Res. https://doi.org/10.1007/s11120-013-9817-2 (2014).

  51. 51.

    Armbruster, U. et al. Arabidopsis Curvature thylakoid1 proteins modify thylakoid architecture by inducing membrane curvature. Plant Cell https://doi.org/10.1105/tpc.113.113118 (2013).

  52. 52.

    Höhner, R. et al. Plastocyanin is the long-range electron carrier between photosystem II and photosystem I in plants. Proc. Natl Acad. Sci. USA https://doi.org/10.1073/pnas.2005832117 (2020).

  53. 53.

    Schreiber, U. Redox changes of ferredoxin, P700, and plastocyanin measured simultaneously in intact leaves. Photosynth. Res. https://doi.org/10.1007/s11120-017-0394-7 (2017).

  54. 54.

    Kirchhoff, H. et al. Dynamic control of protein diffusion within the granal thylakoid lumen. Proc. Natl Acad. Sci. USA https://doi.org/10.1073/pnas.1104141109 (2011).

  55. 55.

    Joliot, P. & Joliot, A. Electron transfer between the two photosystems. II. Equilibrium constants. Biochim Biophys. Acta Bioenerg. https://doi.org/10.1016/0005-2728(84)90016-1 (1984).

  56. 56.

    Kirchhoff, H., Horstmann, S. & Weis, E. Control of the photosynthetic electron transport by PQ diffusion microdomains in thylakoids of higher plants. Biochim. Biophys. Acta Bioenerg. 1459, 148–168 (2000).

    CAS  Article  Google Scholar 

  57. 57.

    Kirchhoff, H., Mukherjee, U. & Galla, H. J. Molecular architecture of the thylakoid membrane: lipid diffusion space for plastoquinone. Biochemistry https://doi.org/10.1021/bi011650y (2002).

  58. 58.

    Yamamoto, H. & Shikanai, T. PGR5-dependent cyclic electron flow protects photosystem I under fluctuating light at donor and acceptor sides. Plant Physiol. 179, 588–600 (2019).

    CAS  Article  Google Scholar 

  59. 59.

    Barbato, R. et al. Higher order photoprotection mutants reveal the importance of ΔpH-dependent photosynthesis-control in preventing light induced damage to both photosystem II and photosystem I. Sci. Rep. 10, 6770 (2020).

    CAS  Article  Google Scholar 

  60. 60.

    Kou, J., Takahashi, S., Fan, D. Y., Badger, M. R. & Chow, W. S. Partially dissecting the steady-state electron fluxes in photosystem I in wild-type and pgr5 and ndh mutants of Arabidopsis. Front. Plant Sci. https://doi.org/10.3389/fpls.2015.00758 (2015).

  61. 61.

    Kadota, K. et al. Oxidation of P700 induces alternative electron flow in photosystem I in wheat leaves. Plants https://doi.org/10.3390/plants8060152 (2019).

  62. 62.

    Nawrocki, W. J. et al. Maximal cyclic electron flow rate is independent of PGRL1 in Chlamydomonas. Biochim. Biophys. Acta Bioenerg. 1860, 425–432 (2019).

    CAS  Article  Google Scholar 

  63. 63.

    Joliot, P. & Alric, J. Inhibition of CO2 fixation by iodoacetamide stimulates cyclic electron flow and non-photochemical quenching upon far-red illumination. Photosynth. Res. https://doi.org/10.1007/s11120-013-9826-1 (2013).

  64. 64.

    Nishio, J. N. & Whitmarsh, J. Dissipation of the proton electrochemical potential in intact chloroplasts. Plant Physiol. https://doi.org/10.1104/pp.101.1.89 (1993).

  65. 65.

    Joliot, P., Lavergne, J. & Béal, D. Plastoquinone compartmentation in chloroplasts. I. Evidence for domains with different rates of photo-reduction. Biochim. Biophys. Acta Bioenerg. 1101, 1–12 (1992).

    CAS  Article  Google Scholar 

  66. 66.

    Johnson, M. P., Vasilev, C., Olsen, J. D. & Hunter, C. N. Nanodomains of cytochrome b6f and photosystem II complexes in spinach grana thylakoid membranes. Plant Cell 26, 3051–3061 (2014).

    CAS  Article  Google Scholar 

  67. 67.

    Schöttler, M. A. & Tóth, S. Z. Photosynthetic complex stoichiometry dynamics in higher plants: environmental acclimation and photosynthetic flux control. Front. Plant Sci. 5, 188 (2014).

    PubMed  PubMed Central  Google Scholar 

  68. 68.

    Pesaresi, P. et al. Mutants, overexpressors, and interactors of Arabidopsis plastocyanin isoforms: revised roles of plastocyanin in photosynthetic electron flow and thylakoid redox state. Mol. Plant https://doi.org/10.1093/mp/ssn041 (2009).

  69. 69.

    Tiwari, A. et al. Photodamage of iron-sulphur clusters in photosystem I induces non-photochemical energy dissipation. Nat. Plants https://doi.org/10.1038/NPLANTS.2016.35 (2016).

  70. 70.

    Takagi, D. & Miyake, C. Proton gradient regulation 5 supports linear electron flow to oxidize photosystem I. Physiol. Plant. https://doi.org/10.1111/ppl.12723 (2018).

  71. 71.

    Foyer, C. H., Neukermans, J., Queval, G., Noctor, G. & Harbinson, J. Photosynthetic control of electron transport and the regulation of gene expression. J. Exp. Bot. 63, 1637–1661 (2012).

    CAS  Article  Google Scholar 

  72. 72.

    Kramer, D. M. & Evans, J. R. The importance of energy balance in improving photosynthetic productivity. Plant Physiol. 155, 70–78 (2011).

    CAS  Article  Google Scholar 

  73. 73.

    Munekage, Y. et al. Cyclic electron flow around photosystem I is essential for photosynthesis. Nature 429, 579–582 (2004).

    CAS  Article  Google Scholar 

  74. 74.

    Giersch, C. et al. Energy charge, phosphorylation potential and proton motive force in chloroplasts. Biochim. Biophys. Acta Bioenerg. https://doi.org/10.1016/0005-2728(80)90146-2 (1980).

  75. 75.

    Backhausen, J. E., Kitzmann, C., Horton, P. & Scheibe, R. Electron acceptors in isolated intact spinach chloroplasts act hierarchically to prevent over-reduction and competition for electrons. Photosynth. Res. https://doi.org/10.1023/A:1026523809147 (2000).

  76. 76.

    Slovacek, R. E., Mills, J. D. & Hind, G. The function of cyclic electron transport in photosynthesis. FEBS Lett. https://doi.org/10.1016/0014-5793(78)80136-7 (1978).

  77. 77.

    Buchert, F., Mosebach, L., Gäbelein, P. & Hippler, M. PGR5 is required for efficient Q cycle in the cytochrome b6f complex during cyclic electron flow. Biochem. J. 477, 1631–1650 (2020).

    CAS  Article  Google Scholar 

  78. 78.

    Mekala, N. R., Suorsa, M., Rantala, M., Aro, E. M. & Tikkanen, M. Plants actively avoid state transitions upon changes in light intensity: role of light-harvesting complex II protein dephosphorylation in high light. Plant Physiol. 168, 721–734 (2015).

    CAS  Article  Google Scholar 

  79. 79.

    Engel, B. D. et al. Native architecture of the Chlamydomonas chloroplast revealed by in situ cryo-electron tomography. eLife https://doi.org/10.7554/eLife.04889 (2015).

  80. 80.

    Iwai, M. et al. Isolation of the elusive supercomplex that drives cyclic electron flow in photosynthesis. Nature 464, 1210–1213 (2010).

    CAS  Article  Google Scholar 

  81. 81.

    Järvi, S., Suorsa, M., Paakkarinen, V. & Aro, E. M. Optimized native gel systems for separation of thylakoid protein complexes: novel super- and mega-complexes. Biochem. J. https://doi.org/10.1042/BJ20102155 (2011).

  82. 82.

    Melis, A. Kinetic analysis of P-700 photoconversion: effect of secondary electron donation and plastocyanin inhibition. Arch. Biochem. Biophys. https://doi.org/10.1016/0003-9861(82)90535-5 (1982).

  83. 83.

    Metzger, S. U., Cramer, W. A. & Whitmarsh, J. Critical analysis of the extinction coefficient of chloroplasty cytochrome f. Biochim. Biophys. Acta Bioenerg. https://doi.org/10.1016/S0005-2728(96)00164-8 (1997).

  84. 84.

    Fristedt, R. et al. Phosphorylation of photosystem II controls functional macroscopic folding of photosynthetic membranes in Arabidopsis. Plant Cell https://doi.org/10.1105/tpc.109.069435 (2009).

  85. 85.

    Hepworth, C., Doheny-Adams, T., Hunt, L., Cameron, D. D. & Gray, J. E. Manipulating stomatal density enhances drought tolerance without deleterious effect on nutrient uptake. New Phytol. https://doi.org/10.1111/nph.13598 (2015).

  86. 86.

    Maxwell, K. & Johnson, G. N. Chlorophyll fluorescence—a practical guide. J. Exp. Bot. https://doi.org/10.1093/jxb/51.345.659 (2000).

  87. 87.

    Huang, W., Suorsa, M. & Zhang, S. B. In vivo regulation of thylakoid proton motive force in immature leaves. Photosynth. Res. https://doi.org/10.1007/s11120-018-0565-1 (2018).

  88. 88.

    Malkin, S., Armond, P. A., Mooney, H. A. & Fork, D. C. Photosystem II photosynthetic unit sizes from fluorescence induction in leaves. Plant Physiol. https://doi.org/10.1104/pp.67.3.570 (1981).

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Acknowledgements

We thank D. Leister (LMU Munich) and M. Pribil (Copenhagen Plant Science Centre) for providing seeds of the psal, curt1abcd, oeCURT1A and tap38 lines and L. Eichacker (University of Stavenger) for providing seeds of stn7. C. Hill (University of Sheffield) is acknowledged for assistance with the EM. M.P.J. acknowledges funding from the Leverhulme Trust grant nos. RPG-2016-161 and RPG-2019-045 and the BBSRC White Rose DTP for a studentship to T.Z.E.M. (BB/M011151/1). The SIM imaging was performed at the University of Sheffield Wolfson Light Microscopy Facility and was partly funded by MRC grant no. MR/K015753/1.

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M.P.J. and S.C. designed the study. C.H., W.H.J.W., T.Z.E.M. and M.S.P. performed the research. C.H., W.H.J.W., T.Z.E.M. and M.P.J. analysed the data. M.P.J., C.H., W.H.J.W., T.Z.E.M., S.C. and M.S.P. wrote the paper.

Corresponding author

Correspondence to Matthew P. Johnson.

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The authors declare no competing interests

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Peer review information Nature Plants thanks Toshiharu Shikanai and the other, anonymous, reviewers for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 PSI and PSII functional antenna size in wild-type and mutant leaves determined by absorption/chlorophyll fluorescence spectroscopy.

PSI antenna size was calculated using P700+ formation kinetics in leaves treated for 1 hour under low light (LL, 125 µmol photons m−2 s−1) or high light (HL, 1150 µmol photons m−2 s−1) followed by infiltration with 30 μM DCMU and 1 mM methyl viologen to induce a donor-limited state around PSI. PSII antenna size was calculated from chlorophyll fluorescence induction curves on LL and HL treated leaves, then infiltrated with 30 μM DCMU. The light intensity used was 12 μmol photons m−2 s−1. P700 traces were fitted with single exponential functions; the tabulated data is the average of four traces per sample. Antenna size was calculated as (WT LL t1/2÷ sample t1/2) × 100 and is expressed as a mean percentage of wild-type LL ± SD. Variable chlorophyll fluorescence was used to calculate the PSII antenna size. The letters a-d represent significant differences calculated using one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test, a-b PSI P=<0.0001, a-c P=<0.0001, a-d P=<0.0001, b-c P=<0.0001, b-d P=<0.0001, c-d P=<0.0001.

Extended Data Fig. 2 Stomatal characterisation of Arabidopsis mutants used in this study.

a, Stomatal density in leaves; mean ± SD is shown for each sample. b, Stomatal conductance in low light (LL, 125 µmol photons m−2 s−1) treated leaves; mean ± SD is shown for each sample. c, Stomatal conductance in high light (HL, 1150 µmol photons m−2 s−1) treated leaves; n (separate plants analysed) = 5-7 for each sample, mean ± SD is shown for each sample. All differences were found to be non-significant by one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test.

Extended Data Fig. 3 Relative levels of cytb6f and PSI complexes and ratios of Pc:PSI and Fd:PSI.

Data determined by spectroscopy on thylakoids from each sample ± SD (n (separate plants analysed) = 3 for each sample); The letters a-b represent significant differences calculated using one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test, a-b PSI P=<0.0001, a-b PSI:Pc P=<0.0001.

Extended Data Fig. 4 Fitting procedure for dark interval relaxation kinetics (DIRK) curves and initial rates of P700 and Pc reduction and Fd oxidation.

a, Half-time obtained from an example DIRK curve for P700+ reduction via a single exponential fit. The fitting residuals are shown underneath. b, The initial rate was calculated using linear fit of the DIRK data from the 3-8 ms window in a.

Extended Data Fig. 5 Investigating possible causes of defective electron transfer regulation.

a, Light-intensity dependence of proton motive force partitioning into ΔpH and ΔΨ according the method of Kramer and Sacksteder, (2000). n = n (separate plants analysed) = 3 for each sample; mean ± SD is shown for each point. b, Equilibrium plot of P700/P700+ versus Pc/Pc+ from dark interval relaxation kinetics following low light + far-red light treatment shown in Fig. 3b (low light = 125 µmol photons m−2 s−1, far-red = 740 nm, 50 μmol photons m−2 s-1). Apparent equilibrium constants (Kapp) calculated from a linear fit of the slope. c, Light-intensity dependence of proton conductivity (gH+). n (separate plants analysed) = 3 for each sample; mean ± SD is shown for each point.

Extended Data Fig. 6 Thin-section EM of membrane architectural changes.

a, EM image showing a chloroplast from WT leaf treated for 1 hour under low light (LL, 125 µmol photons m−2 s−1), b, EM image showing a chloroplast from WT leaf treated for 1 hour under high light (HL, 1150 µmol photons m−2 s−1). Scale bars 200 nm. c, Cross-section through a grana stack with repeat lumen width labelled. d, lumen width of grana layers in LL (n (grana analysed) = 21) and HL (n (grana analysed) = 21) adapted WT leaves. e, lumen width of grana layers in HL adapted WT (n (grana analysed) = 21), tap38 (n (grana analysed) = 19) and stn7 (n (grana analysed)= 23) leaves. All differences were found non-significant by one-way analysis of variance (ANOVA). Two independent sets of micrographs were obtained with similar results.

Extended Data Fig. 7 Analysis of cytb6f distribution in thylakoid membranes from WT and mutant Arabidopsis plants.

Spectroscopic analysis of chlorophyll distribution between fractions and cytochrome f/chlorophyll content (cyt f/ chl) was performed on thylakoids isolated from leaves treated for 1 hour under low light (LL, 125 µmol photons m−2 s−1) or high light (HL, 1150 µmol photons m−2 s−1). The letters a-b represent significant differences calculated using one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test, a-b PSI P=<0.0001.

Extended Data Fig. 8 PSI function and Infiltration control experiments.

a, % functional PSI remaining after 2 hours HL (1150 µmol photons m−2 s−1) treatment. Total functional PSI calculated by amplitude of the ECS a-phase triggered by a 50 μs 635 nm pulse upon infiltration of dark-adapted leaves with 30 μM DCMU. This was compared to HL treated leaves that were subsequently infiltrated with DCMU and % remaining calculated. n (separate plants) = 5 for each, mean ± S.D. The letters a-b represent significant differences calculated using one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test, a-b PSI P=<0.0001. b, Proton conductivity (gH+) calculated upon before (dark) and after 15 minutes treatment with FR light (740 nm, 255 µmol photons m−2 s−1) on leaves infiltrated with 20 mM Hepes pH 7.5, 150 mM sorbitol, 50 mM NaCl, 4 mM iodoacetamide (IA). n (separate plants) = 3, mean ± SD is shown for each condition. c, Rapidly-reversible NPQ (qE) following 10 minutes HL (1150 µmol photons m−2 s−1) illumination of leaves infiltrated with either 20 mM Hepes pH 7.5, 150 mM sorbitol, 50 mM NaCl (control) or the same buffer swapping NaCl for 50 mM NaNO2. n (separate plants) = 5 for each sample; mean ± SD is shown for each condition. The letters a-b represent significant differences calculated using one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test, a-b PSI P=<0.0001.

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Hepworth, C., Wood, W.H.J., Emrich-Mills, T.Z. et al. Dynamic thylakoid stacking and state transitions work synergistically to avoid acceptor-side limitation of photosystem I. Nat. Plants 7, 87–98 (2021). https://doi.org/10.1038/s41477-020-00828-3

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