Abstract
Chloroplast-actin (cp-actin) filaments are crucial for light-induced chloroplast movement, and appear in the front region of moving chloroplasts when visualized using GFP-mouse Talin. They are short and thick, exist between a chloroplast and the plasma membrane, and move actively and rapidly compared to cytoplasmic long actin filaments that run through a cell. The average period during which a cp-actin filament was observed at the same position was less than 0.5 s. The average lengths of the cp-actin filaments calculated from those at the front region of the moving chloroplast and those around the chloroplast periphery after stopping the movement were almost the same, approximately 0.8 µm. Each cp-actin filament is shown as a dotted line consisting of 4–5 dots. The vector sum of cp-actin filaments in a moving chloroplast is parallel to the moving direction of the chloroplast, suggesting that the direction of chloroplast movement is regulated by the vector sum of cp-actin filaments. However, once the chloroplasts stopped moving, the vector sum of the cp-actin filaments around the chloroplast periphery was close to zero, indicating that the direction of movement was undecided. To determine the precise structure of cp-actin filaments under electron microscopy, Arabidopsis leaves and fern Adiantum capillus-veneris gametophytes were frozen using a high-pressure freezer, and observed under electron microscopy. However, no bundled microfilaments were found, suggesting that the cp-actin filaments were unstable even under high-pressure freezing.
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Acknowledgements
We thank Dr. Saku T. Kijima, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Ibaraki 305-8562, Japan, for the dormitory arrangement at AIST.
This work was supported in part by Grants-in-Aid for scientific research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT)/the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers 20227001, 23120523, 25120721, 25251033, and 16K14758 (to M.W) and by the Ohsumi Frontier Science Foundation (to M.W.).
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MW conceived the study, MW and YM designed the study, MW, TH and KK performed the live imaging experiments, MW, TH, YM, TN, NM, SS, YN and AM performed the electron microscopy experiments, MW and YM analyzed the data, MW and YM wrote the manuscript.
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Wada, M., Higa, T., Katoh, K. et al. Chloroplast-actin filaments decide the direction of chloroplast avoidance movement under strong light in Arabidopsis thaliana. J Plant Res (2024). https://doi.org/10.1007/s10265-024-01540-5
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DOI: https://doi.org/10.1007/s10265-024-01540-5