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Thylakoid membrane stacking controls electron transport mode during the dark-to-light transition by adjusting the distances between PSI and PSII

Nature Plants volume 10, pages 512–524 (2024)Cite this article

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Abstract

The balance between linear electron transport (LET) and cyclic electron transport (CET) plays an essential role in plant adaptation and protection against photo-induced damage. This balance is largely maintained by phosphorylation-driven alterations in the PSII–LHCII assembly and thylakoid membrane stacking. During the dark-to-light transition, plants shift this balance from CET, which prevails to prevent overreduction of the electron transport chain and consequent photo-induced damage, towards LET, which enables efficient CO2 assimilation and biomass production. Using freeze-fracture cryo-scanning electron microscopy and transmission electron microscopy of Arabidopsis leaves, we reveal unique membrane regions possessing characteristics of both stacked and unstacked regions of the thylakoid network that form during this transition. A notable consequence of the morphological attributes of these regions, which we refer to as ‘stacked thylakoid doublets’, is an overall increase in the proximity and connectivity of the two photosystems (PSI and PSII) that drive LET. This, in turn, reduces diffusion distances and barriers for the mobile carriers that transfer electrons between the two PSs, thereby maximizing LET and optimizing the plant’s ability to utilize light energy. The mechanics described here for the shift between CET and LET during the dark-to-light transition are probably also used during chromatic adaptation mediated by state transitions.

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Fig. 1: Morphometric characterization of Arabidopsis WT, stn7 stn8 and pph1 pbcp plants.
Fig. 2: Density of PSII complexes in stacked and unstacked thylakoids of dark- and light-adapted plants.
Fig. 3: Segmentation and quantification of thylakoid domains.
Fig. 4: PSII fraction residing in proximity to PSI.
Fig. 5: Photosynthetic performance assessed by chlorophyll a fluorescence recordings.
Fig. 6: A model for the dark-to-light transition.

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Data availability

The data are available in the article, Extended Data Figs. 1–3 and Extended Data Tables 1 and 2. The images are deposited at figshare (https://doi.org/10.6084/m9.figshare.24942420). Source data are provided with this paper.

Code availability

The MATLAB code is available in Supplementary Code 1.

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Acknowledgements

This work is dedicated to the memory of Eyal Shimoni, who passed away in July 2023. This work was supported by grants from the Israel Science Foundation (no. 1082/17 to Z.R. and R.N.; no. 1377/18 to D.C.) and the National Science Foundation United States–Israel Binational Science Foundation Molecular and Cellular Biosciences Program (no. 1616982 to H.K.; no. 2019695 to Z.R. and R.N.; no. 2015839 to Z.R.; no. 1953570 to H.K.).

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  1. Deceased: Eyal Shimoni.

Authors and Affiliations

  1. Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel

    Yuval Garty, Yuval Bussi, Reinat Nevo & Ziv Reich

  2. Department of Chemical Research Support, Weizmann Institute of Science, Rehovot, Israel

    Smadar Levin-Zaidman & Eyal Shimoni

  3. Institute of Biological Chemistry, Washington State University, Pullman, WA, USA

    Helmut Kirchhoff

  4. Institute of Plant Sciences, Agricultural Research Organization, Volcani Institute, Rishon LeZion, Israel

    Dana Charuvi

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Contributions

Z.R., H.K. and R.N. designed the research. Y.G., S.L.-Z. and E.S. performed the experiments. Y.G., Y.B., D.C. and R.N. analysed the data. Y.B., D.C., Z.R., H.K. and R.N. wrote the manuscript.

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Correspondence to Reinat Nevo or Ziv Reich.

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Extended data

Extended Data Fig. 1 Representative cryo-SEM images of freeze-fractured leaf samples from dark- and light-adapted plants.

The exoplasmic fracture faces of unstacked membranes (EFu) are outlined. Scale bars: 100 nm. Images shown are representative images of the experiment described in Fig. 2.

Extended Data Fig. 2 Representative thin-section TEM images of chloroplasts (means ± SE) and a box plot of grana widths from dark- and light-adapted leaves of WT and of the stn7/stn8 and pph1/pbcp mutants.

N = number of grana probed per genotype and condition: WT - 291 (D), 232 (L); stn7/stn8 - 160 (D), 126 (L); pph1/pbcp - 140 (D), 140 (L). For each genotype and condition, samples were obtained from two different plants. In the box plots, the box indicates the interquartile area, whiskers are drawn down to the 5th percentile and up to the 95th, with small black squares in the middle representing the means. Scale bars: 500 nm.

Extended Data Fig. 3 Simulation of PSII to PSI nearest neighbor distance using empirical (PSII) and calculated (PSI) densities in unstacked membranes in WT.

For the obtained PSI:PSII ratios, of 5.1 for dark (D) and 3.1 for light (L) (gray lines), the analysis showed that the PSII-PSI nearest neighbor distances were ˂2 nm. For the simulation, PSI and PSII complexes were represented by disks with areas approximated from their PDB structures (PSI [2WSC]; PSII [7OUI]). PSII particles were randomly placed in a 1 µm2 grid with the densities observed in dark (D) and light (L) conditions (522 particles/µm2 and 773 particles/µm2, respectively, see Fig. 2). PSI particles were randomly placed in unoccupied positions (not allowing for particle overlap) until either no more particles could be added randomly or the PSI density reached our estimated values (2685 particles/µm2 [D] or 2410 particles/µm2 [L])*. Using the PSII and PSI densities resulted in PSI:PSII ratios of 5.1 for D (dark grey) and 3.1 for L (light grey). The simulation was carried out 100 times for different PSI:PSII ratios, three examples for ratios of 1:1; 2:1; 3:1 in the dark condition are shown with PSII colored in green and PSI in magenta. The plot depicts the PSII-PSI nearest neighbor distance (from particle edge to edge) for D (blue) and L (red), with points representing the mean of the means and error bars showing the mean of the standard deviations for the distances in the 100 replicates (N = 100; Data are presented as means of means ± means of SDs). For both D and L, at the calculated PSI:PSII ratios (5.1 [D] and 3.1 [L]) when the maximum PSI particles were placed in the grid randomly, the extrapolated PSII-PSI nearest neighbor distances were ˂2 nm. The simulation (Garty_at_al_Supplementary_information_2.m) was carried out using MATLAB version: 9.13.0 (R2022b), Natick, Massachusetts: The MathWorks Inc.; 2022. *Values of PSI densities were estimated using a PSII:PSI ratio of 1.3 for the whole thylakoid fraction89. We then calculated the ratios of stacked/unstacked membranes, from the observed thylakoid fractions (Fig. 3f), as \(\frac{\frac{1}{2}{f}_{GSL}+{f}_{G}}{\frac{1}{2}{f}_{GSL}+{f}_{SL}}\). The values obtained, 1.94 in dark and 1.56 in light, were used to calculate the PSI density, which was similar to published AFM data29.

Extended Data Table 1 Maximum quantum yield of PSII (Fv/Fm)*
Full size table
Extended Data Table 2 Thylakoid length* in dark- and light-adapted plants
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Supplementary information

Reporting Summary

Supplementary Code 1

Code for Extended Data Fig. 3.

Source data

Source Data Fig. 1

Statistical source data for Figs. 1–5.

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Garty, Y., Bussi, Y., Levin-Zaidman, S. et al. Thylakoid membrane stacking controls electron transport mode during the dark-to-light transition by adjusting the distances between PSI and PSII. Nat. Plants 10, 512–524 (2024). https://doi.org/10.1038/s41477-024-01628-9

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