Ute Armbruster for providing the seeds of the Ler0 curt1a-1 mutant. This work was supported by the Dutch Organisation for Scientific Research (NWO) via a Vidi grant no. VI.Vidi 192.042 (E.W.) and by Wageningen Graduates Schools through a PhD grant (J.B.). Data availability The data underlying this publication can be accessed at https://doi.org/10.4121/75fa3c66-8505-4d6a-9bd9-16973e5ca885.References [1] R.E. Blankenship, Molecular Mechanisms of Photosynthesis, John Wiley & Sons, 2021, https://doi.org/10.1002/9780470758472. [2] H. Kirchhoff, Chloroplast ultrastructure in plants, New Phytol. 223 (2) (2019) 565–574, https://doi.org/10.1111/nph.15730. [3] H. Kirchhoff, C. Hall, M. Wood, M. Herbst
Open resource ↗10.4121/75fa3c66-8505-4d6a-9bd9-16973e5ca885 · pdf-raw-page:9 lines:1-68Unverified paper record
Expansion microscopy reveals thylakoid organisation alterations due to genetic mutations and far-red light acclimation.
Biochimica et biophysica acta. Bioenergetics · 13 Mar 2025 · 10.1016/j.bbabio.2025.149552
Abstract
The thylakoid membrane is the site of the light-dependent reactions of photosynthesis. It is a continuous membrane, folded into grana stacks and the interconnecting stroma lamellae. The CURVATURE THYLAKOID1 (CURT1) protein family is involved in the folding of the membrane into the grana stacks. The thylakoid membrane remodels its architecture in response to light conditions, but its 3D organisation and dynamics remain incompletely understood. To resolve these details, an imaging technique is needed that provides high-resolution 3D images in a high-throughput manner. Recently, we have used expansion microscopy, a technique that meets these criteria, to visualise the thylakoid membrane isolated from spinach. Here, we show that this protocol can also be used to visualise enveloped spinach chloroplasts. Additionally, we present an improved protocol for resolving the thylakoid structure of Arabidopsis thaliana. Using this protocol, we show the changes in thylakoid architecture in response to long-term far-red light acclimation and due to knocking out CURT1A. We show that far-red light acclimation results in higher grana stacks that are packed closer together. In addition, the distance between stroma lamellae, which are wrapped around the grana, decreases. In the curt1a mutant, grana have an increased diameter and height, and the distance between grana is increased. Interestingly, in this mutant, the stroma lamellae occasionally approach the grana stacks from the top. These observations show the potential of expansion microscopy to study the thylakoid membrane architecture.
Plant phenotyping relevance
植物のチラコイド膜構造を高解像度3D画像で取得する拡大顕微鏡法の改良・適用が中心であり、膜構造という植物形態形質を測定しているため。
abstractTo resolve these details, an imaging technique is needed that provides high-resolution 3D images in a high-throughput manner.
abstractAdditionally, we present an improved protocol for resolving the thylakoid structure of Arabidopsis thaliana.
abstractThese observations show the potential of expansion microscopy to study the thylakoid membrane architecture.
Code and data availability
The article states that the data underlying the publication (expansion microscopy imaging/measurements of thylakoid architecture) are publicly available in the 4TU Research Data repository via the DOI 10.4121/75fa3c66-8505-4d6a-9bd9-16973e5ca885. This is a paper-specific, publicly accessible data deposit with an author
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