Abstract
Chloroplast movement is regulated by dynamic reorganization of the actin cytoskeleton. To gain insight into the function of ACT2 in chloroplast movement, we examined the effect of the act2-3 mutation, in which a T-DNA is inserted at the second exon of the ACT2 gene, and investigated a transgenic Arabidopsis plant expressing the GFP-ACT2 fusion protein. Chloroplast movement in the act2-3 mutant was retarded, especially during the avoidance response, compared with that in the WT. We further verified the physiological response of transgenic Arabidopsis expressing the GFP-ACT2 fusion gene with a 6xGSS linker sequence, which was recently reported to enable the direct visualization of actin filaments in a transient expression system. Although the expression of GFP-ACT2 had highly detrimental effects on the growth and development of transgenic Arabidopsis plants, GFP-ACT2 was incorporated into chloroplast actin (cp-actin) filaments instead of cortical actin filaments. The analyses of the act2 mutant and GFP-ACT2 transgenic plants suggest that ACT2 functions in chloroplast movement as a component of cp-actin filaments and that cp-actin filaments are differently regulated compared to cortical actin filaments.
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This work was supported by a research grant from Kongju National University in 2016.
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S-GK designed and supervised the study. J-YK, JA, and HB performed the experiments. J-YK, JA, and S-GK analyzed the data. J-YK, JA, and S-GK wrote the manuscript. MW provided experimental tools. S-GK revised the manuscript. All the authors read and approved the manuscript.
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Supplementary file2 Supplemental Movie 1. Chloroplast photorelocation movement induced by microbeam irradiation in a WT cell (AVI 2164 kb)
Supplementary file3 Supplemental Movie 2. Chloroplast photorelocation movement induced by microbeam irradiation in an act2-3 mutant cell (AVI 2078 kb)
Supplementary file4 Supplemental Movie 3. Cp-actin filament dynamics during the strong light-induced avoidance response in GFP-TALIN cells (AVI 2156 kb)
Supplementary file5 Supplemental Movie 4. Cp-actin filament dynamics during the strong light-induced avoidance response in GFP-ACT2/WT cells (AVI 2156 kb)
Supplementary file6 Supplemental movie 5. Cp-actin filament dynamics during the strong light-induced avoidance response in GFP-ACT2/act2-3 cells (AVI 2156 kb)
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Kim, Jy., Ahn, J., Bong, H. et al. ACTIN2 Functions in Chloroplast Photorelocation Movement in Arabidopsis thaliana. J. Plant Biol. 63, 379–389 (2020). https://doi.org/10.1007/s12374-020-09262-6
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DOI: https://doi.org/10.1007/s12374-020-09262-6