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Chloroplast actin filaments organize meshwork on the photorelocated chloroplasts in the moss Physcomitrella patens

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Abstract

Cytoskeleton dynamics during phototropin-dependent chloroplast photorelocation movement was analyzed in protonemal cells of actin- and microtubule-visualized lines of Physcomitrella patens expressing GFP- or tdTomato-talin and GFP-tubulin. Using newly developed epi- and trans-microbeam irradiation systems that permit fluorescence observation of the cell under blue microbeam irradiation inducing chloroplast relocation, it was revealed that meshwork of actin filaments formed at the chloroplast-accumulating area both in the avoidance and accumulation movements. The structure disappeared soon when blue microbeam was turned off, and it was not induced under red microbeam irradiation that did not evoke chloroplast relocation movement. In contrast, no apparent change in microtubule organization was detected during the movements. The actin meshwork was composed of short actin filaments distinct from the cytoplasmic long actin cables and was present between the chloroplasts and plasma membrane. The short actin filaments emerged from around the chloroplast periphery towards the center of chloroplast. Showing highly dynamic behavior, the chloroplast actin filaments (cp-actin filaments) were rapidly organized into meshwork on the chloroplast surface facing plasma membrane. The actin filament configuration on a chloroplast led to the formation of actin meshwork area in the cell as the chloroplasts arrived at and occupied the area. After establishment of the meshwork, cp-actin filaments were still highly dynamic, showing appearance, disappearance, severing and bundling of filaments. These results indicate that the cp-actin filaments have significant roles in the chloroplast movement and positioning in the cell.

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Abbreviations

BDM:

2,3-Butanedion monoxime

cp-actin filament:

Chloroplast actin filament

References

  • Anielska-Mazur A, Bernas T, Gabrys H (2009) In vivo reorganization of the actin cytoskeleton in leaves of Nicotiana tabacum L. transformed with plastin-GFP. Correlation with light-activated chloroplast responses. BMC Plant Biol 9:64

    Article  PubMed  Google Scholar 

  • Dong X-J, Takagi S, Nagai R (1995) Regulation of the orientation movement of chloroplasts in epidermal cells of Vallisneria. Protoplasma 195:18–24

    Article  Google Scholar 

  • Dong X-J, Nagai R, Takagi S (1998) Microfilaments anchor chloroplasts along the outer periclinal wall in Vallisneria epidermal cells through cooperation of PFR and photosynthesis. Plant Cell Physiol 36:1299–1306

    Google Scholar 

  • Gabrys H (2004) Blue light-induced orientation movements of chloroplasts in higher plants: recent progress in the study of their mechanisms. Acta Physiol Plant 26:473–478

    Article  Google Scholar 

  • Higaki T, Sano T, Hasezawa S (2007) Actin microfilament dynamics and actin side-binding proteins in plants. Curr Opin Plant Biol 10:549–556

    Article  CAS  PubMed  Google Scholar 

  • Hiwatashi Y, Obara M, Sato Y, Fujita T, Murata T, Hasebe M (2008) Kinesins are indispensable for interdigitation of phragmoplast microtubules in the moss Physcomitrella patens. Plant Cell 20:3094–3106

    Article  CAS  PubMed  Google Scholar 

  • Hussey PJ, Allwood EG, Smertenko AP (2002) Actin-binding proteins in the Arabidopsis genome database: properties of functionally distinct plant actin-depolymerizing factors/cofilins. Phil Trans Roy Soc Lond B 357:791–798

    Article  CAS  Google Scholar 

  • Kadota A, Wada M (1989) Photoinduction of circular F-actin on chloroplast in a fern protonemal cell. Protoplasma 151:171–174

    Article  Google Scholar 

  • Kadota A, Wada M (1992) Photoinduction of formation of circular structures by microfilaments on chloroplasts during intracellular orientation in protonemal cells of the fern Adiantum capilus-veneris. Protoplasma 167:97–107

    Article  Google Scholar 

  • Kadota A, Sato Y, Wada M (2000) Intracellular chloroplast photorelocation in the moss Physcomitrella patens is mediated by phytochrome as well as by a blue-light receptor. Planta 210:932–937

    Article  CAS  PubMed  Google Scholar 

  • Kadota A, Yamada N, Suetsugu N, Hirose M, Saito C, Shoda K, Ichikawa S, Kagawa T, Nakano A, Wada M (2009) Short actin-based mechanism for light-directed chloroplast movement in Arabidopsis. Proc Natl Acad Sci USA 106:13106–13111

    Article  CAS  PubMed  Google Scholar 

  • Kagawa T, Sakai T, Suetsugu N, Oikawa K, Ishiguro S, Kato T, Tabata S, Okada K, Wada M (2001) Arabidopsis NPL1: a phototropin homolog controlling the chloroplast high-light avoidance response. Science 291:2138–2141

    Article  CAS  PubMed  Google Scholar 

  • Kagawa T, Kasahara M, Abe T, Yoshida S, Wada M (2004) Function analysis of phototropin2 using fern mutants deficient in blue light-induced chloroplast avoidance movement. Plant Cell Physiol 45:416–426

    Article  CAS  PubMed  Google Scholar 

  • Kandasamy MK, Meagher RB (1999) Actin-organelle interaction: association with chloroplast in Arabidopsis leaf mesophyll cells. Cell Motil Cytoskeleton 44:110–118

    Article  CAS  PubMed  Google Scholar 

  • Kasahara M, Kagawa T, Sato Y, Kiyosue T, Wada M (2004) Phototropins mediate blue and red light-induced chloroplast movements in Physcomitrella patens. Plant Physiol 135:1388–1397

    Article  CAS  PubMed  Google Scholar 

  • Kawai H, Kanegae T, Christensen S, Kiyosue T, Sato Y, Imaizumi T, Kadota A, Wada M (2003) Responses of ferns to red light are mediated by an unconventional photoreceptor. Nature 421:287–290

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi H, Fukuda H, Shibaoka H (1987) Reorganization of actin filaments associated with the differentiation of tracheary elements in Zinnia mesophyll cells. Protoplasma 138:69–71

    Article  Google Scholar 

  • Kost B, Spielhofer P, Chua NH (1998) A GFP-mouse talin fusion protein labels plant actin filaments in vivo and visualizes the actin cytoskeleton in growing pollen tubes. Plant J 16:393–401

    Article  CAS  PubMed  Google Scholar 

  • Kumatani T, Sakurai-Ozato N, Miyawaki N, Yokota E, Shimmen T, Terashima I, Takagi S (2006) Possible association of actin filaments with chloroplasts of spinach mesophyll cells in vivo and in vitro. Protoplasma 229:45–52

    Article  CAS  PubMed  Google Scholar 

  • McElroy D, Zhang W, Cao J, Wu R (1990) Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2:163–171

    Article  CAS  PubMed  Google Scholar 

  • Nishiyama T, Hiwatashi Y, Sakakibara I, Kato M, Hasebe M (2000) Tagged mutagenesis and gene-trap in the moss, Physcomitrella patens by shuttle mutagenesis. DNA Res 7:9–17

  • Nozue K, Kanegae T, Imaizumi T, Fukuda S, Okamoto H, Yeh K-C, Lagarias JC, Wada M (1998) A phytochrome from the fern Adiantum with features of the putative photoreceptor NPH1. Proc Natl Acad Sci USA 95:15826–15830

    Article  CAS  PubMed  Google Scholar 

  • Sakai Y, Takagi S (2005) Reorganized actin filaments anchor chloroplasts along the anticlinal walls of Vallisneria epidermal cells under high-intensity blue light. Planta 221:823–830

    Article  CAS  PubMed  Google Scholar 

  • Sakai T, Kagawa T, Kasahara M, Swartz TE, Christie JM, Briggs WR, Wada M, Okada K (2001) Arabidopsis nph1 and npl1: blue light receptors that mediate both phototropism and chloroplast relocation. Proc Natl Acad Sci USA 98:6969–6974

    Article  CAS  PubMed  Google Scholar 

  • Sato Y, Wada M, Kadota A (2001) Choice of tracks, microtubules and/or actin filaments for chloropalst photo-movement is differentially controlled by phytochrome and a blue light receptor. J Cell Sci 114:269–279

    CAS  PubMed  Google Scholar 

  • Shaner NC, Campbell RE, Steinbach PA, Giepmans BNG, Palmer AE, Tsien RY (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp red fluorescent protein. Nat Biotechnol 22:1567–1572

    Article  CAS  PubMed  Google Scholar 

  • Sheahan MB, Staiger CJ, Rose RJ, McCurdy DW (2004) A green fluorescent protein fusion to actin-binding domain 2 of Arabidopsis fimbrin highlights new features of a dynamic actin cytoskeleton in live plant cells. Plant Physiol 136:3968–3978

    Article  CAS  PubMed  Google Scholar 

  • Shimmen T (2007) The sliding theory of cytoplasmic streaming: fifty years of progress. J Plant Res 120:31–43

    Article  CAS  PubMed  Google Scholar 

  • Shimmen T, Yokota E (2004) Cytoplasmic streaming in plants. Curr Opin Cell Biol 16:68–72

    Article  CAS  PubMed  Google Scholar 

  • Staiger CJ, Blanchoin L (2006) Actin dynamics: old friends with new stories. Curr Opin Plant Biol 9:554–562

    Article  CAS  PubMed  Google Scholar 

  • Staiger CJ, Sheahan MB, Khurana P, Wang X, McCurdy DW, Blanchoin L (2009) Actin filament dynamics are dominated by rapid growth and severing activity in the Arabidopsis cortical array. J Cell Biol 184:269–280

    Article  CAS  PubMed  Google Scholar 

  • Staiger CJ, Poulter NS, Henty JL, Franklin-Tong VE, Blanchoin L (2010) Regulation of actin dynamics by actin-binding proteins in pollen. J Exp Bot 61:1969–1986

    Article  CAS  PubMed  Google Scholar 

  • Suetsugu N, Wada M (2007a) Chloroplast photorelocation movement mediated by phototropin family proteins in green plants. Biol Chem 388:927–935

    Article  CAS  PubMed  Google Scholar 

  • Suetsugu N, Wada M (2007b) Phytochrome-dependent photomovement responses mediated by phototropin family proteins in cryptogam plants. Photochem Photobiol 83:87–93

    CAS  PubMed  Google Scholar 

  • Suetsugu N, Mittmann F, Wagner G, Hughes J, Wada M (2005) A chimeric photoreceptor gene, NEOCHROME, has arisen twice during plant evolution. Proc Natl Acad Sci USA 102:13705–13709

    Article  CAS  PubMed  Google Scholar 

  • Takagi S (2002) Actin-based photo-orientation movement of chloroplasts in plant cells. J Exp Biol 206:1963–1969

    Article  Google Scholar 

  • Thomas C, Tholl S, Moes D, Dieterle M, Papuga J, Moreau F, Steinmetz A (2009) Actin bundling in plants. Cell Motil Cytoskeleton 66:940–957

    Article  CAS  PubMed  Google Scholar 

  • Ueda H, Yokota E, Kutsuna N, Shimada T, Tamura K, Shimmen T, Hasezawa S, Dolja VV, Hara-Nishimura I (2010) Myosin-dependent endoplasmic reticulum motility and F-actin organization in plant cells. Proc Natl Acad Sci USA 107:6894–6899

    Article  CAS  PubMed  Google Scholar 

  • van der Honing HS, Emons AMC, Ketelaar T (2007) Actin based processes that could determine the cytoplasmic architecture of plant cells. Biochim Biophys Acta 1773:604–614

    Article  PubMed  Google Scholar 

  • Vidali L, Rounds CM, Hepler PK, Bezanilla M (2009) Lifeact-mEGFP reveals a dynamic apical f-actin network in tip growing plant cells. PLoS ONE 4:e5744

    Article  PubMed  Google Scholar 

  • Wada M, Suetsugu N (2004) Plant organelle positioning. Curr Opin Plant Biol 7:626–631

    Article  CAS  PubMed  Google Scholar 

  • Wada M, Kagawa T, Sato Y (2003) Chloroplast movement. Annu Rev Plant Biol 54:455–468

    Article  CAS  PubMed  Google Scholar 

  • Wang YS, Motes CM, Mohamalawari DR, Blancaflor EB (2004) Green fluorescent protein fusions to Arabidopsis fimbrin 1 for spatio-temporal imaging of F-actin dynamics in roots. Cell Motil Cytoskeleton 59:79–93

    Article  CAS  PubMed  Google Scholar 

  • Wang YS, Yoo CM, Blancaflor EB (2008) Improved imaging of actin filaments in transgenic Arabidopsis plants expressing a green fluorescent protein fusion to the C- and N-termini of the fimbrin actin-binding domain 2. New Phytol 177:525–536

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the Grant-in-Aid for Scientific Research (19039027, 19570045 and 22570047 to AK; 13139203, 13304061, 16107002 and 20227001 to MW; 20570041 to TK) from the Ministry of Education, Sports, Science and Technology of Japan. Initial studies of this work were done by Ms. M. Nagai and Ms. T. Ashizawa (Tokyo Metropolitan University). We thank Dr. M. Hasebe (NIBB) for the gift of several vector constructs for P. patens transformation, Dr. R. Y. Tsien (UCSD) for the gift of tdTomato cDNA and Mr. N. Yamada (Tokyo Metropolitan University) for the illustration of microbeam irradiation systems.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biological Sciences, Graduate School of Science and Engineering, Tokyo Metropolitan University, Tokyo, 192-0397, Japan

    Hiroko Yamashita, Takeshi Kanegae & Akeo Kadota

  2. Division of Evolutionary Biology, National Institute for Basic Biology, Okazaki, 444-8585, Japan

    Yoshikatsu Sato

  3. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, 305-8577, Japan

    Takatoshi Kagawa

  4. Department of Biology, Faculty of Sciences, Kyushu University, Fukuoka, 812-8581, Japan

    Masamitsu Wada

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  1. Hiroko Yamashita
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  2. Yoshikatsu Sato
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  3. Takeshi Kanegae
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  4. Takatoshi Kagawa
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  5. Masamitsu Wada
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Corresponding author

Correspondence to Akeo Kadota.

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Fig. S1

Scheme of epi-microbeam irradiation system. In addition to the optical path for ordinal transmission observation, the epi-microbeam irradiation system is equipped with two optical paths, one for epifluorescence microscopy and the other for microbeam irradiation with the stimulus light. The system permitted time-lapse recording of fluorescence image at defined intervals in epi-fluorescence mode, as well as continuous microbeam irradiation of cells with defined wavelengths and intensities in epi-microbeam mode excluding the period of fluorescence image acquisition. Switching between the two modes was performed by turning a mirror connecting the two optical systems (PPT 52 kb)

Fig. S2

Scheme of trans-microbeam irradiation system. The trans-microbeam irradiation system is equipped withtwo optical paths, one for epifluorescence microscopy and the other for ordinal transmission microscopy. For trans-microbeam irradiation, slit and interference filter was placed in the latter light path to illuminate the part of the cell with defined wavelengths and intensities. Microbeam irradiation with the stimulus light was performed continuously, being independent from fluorescence image acquisition (PPT 36 kb)

Movie S1

Actin meshwork locates between plasma membrane and chloroplasts. These movies show the Z-scanned fluorescence images of the cell irradiated with a microbeam (10 μm in width) of weak blue light (2.4 W m−2) for 1 h (Movie S1) and with that of strong blue light (96 W m−2) for 1 h thereafter (Movie S2). Note the actin meshwork is seen only on the plasma membrane side of chloroplasts. Parts of these movies were presented in Fig. 1b (MOV 1619 kb)

Movie S2

See caption of Supplementary Movie S1 (MOV 1962 kb)

Movie S3

Reorganization of actin filaments and microtubules during blue light-induced accumulation and avoidance movements of chloroplasts. This movie shows the reorganization of actin filaments and microtubules during chloroplast photomovement. Protonemal cell was irradiated with a microbeam of weak blue light (1 W m−2) for 50 min and then of strong blue light (100 W m−2) for 65 min. Area of microbeam irradiation is indicated with blue lines at the beginning of each treatment. Note the actin meshwork appears on the chloroplasts at microbeam area under weak light irradiation and at both sides of microbeam under strong light irradiation. Selected images of this movie are presented in Fig. 2 (MOV 4599 kb)

Appearance and disappearance of cp-actin filaments on the chloroplast. These movies show the actin meshwork formation on the chloroplasts under microbeam irradiation with strong blue light (Movie S4) and the disappearance of the meshwork in the dark (Movie S5). Cell was treated with both 0.25 mM oryzalin and 25 mM BDM for 2 h and then irradiated with a strong blue light microbeam (20 μm in width, 175 W m−2). An area next to the microbeam (shown in Fig. 7a) was observed. Several images of these movies are presented in Fig. 7b, c (MOV 2975 kb)

See caption of Supplementary Movie S4 (MOV 1009 kb)

Actin dynamics in the meshwork. These movies show the dynamics of actin meshwork. Protonemal cell was irradiated with blue light microbeam (175 W m−2) for 60 min and the dynamics of the actin meshwork formed next to microbeam area (shown in Fig. 8a) were observed every 5 s under microbeam irradiation. Several images of these movies are presented in Fig. 8b (MOV 256 kb)

See caption of Supplementary Movie S6 (MOV 214 kb)

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Yamashita, H., Sato, Y., Kanegae, T. et al. Chloroplast actin filaments organize meshwork on the photorelocated chloroplasts in the moss Physcomitrella patens . Planta 233, 357–368 (2011). https://doi.org/10.1007/s00425-010-1299-2

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