Summary.
The ability of plant cells to dedifferentiate represents an important survival strategy invoked in a range of situations from repair mechanisms following wounding to apomixis. Dedifferentiation requires that somatic cells reprogram and enter the cell division cycle. This in turn necessitates the accurate partitioning of nuclear content and organelles, such as chloroplasts, to daughter cells, thereby ensuring continuity of cellular information systems. The distribution of cytoplasm and its organelle content in mature plant cells is governed by a large, central vacuole, with connections between distant cortical and perinuclear cytoplasmic domains mediated by transvacuolar strands. Here we examined the changes to vacuolar architecture in Arabidopsis thaliana protoplasts expressing a green-fluorescent protein fusion to a δ-tonoplast-intrinsic protein (δTIP). We found that vacuolar architecture became increasingly intricate during protoplast culture with the development of numerous transvacuolar strands. The development of an intricate vacuolar architecture was an actin filament- and not microtubule-dependent process, as is the case in interphase plant cells. Furthermore, we show that myosin is required for this increased complexity of vacuolar architecture and the formation of subcortical actin filament arrays. Despite the likelihood that increased vacuolar invagination would allow better redistribution of cytoplasmic organelles, we found that repositioning of chloroplasts from cortical to perinuclear cytoplasm was not dependent on transvacuolar strands. Our findings indicate that the vacuole is a dynamic entity that develops a complex architecture before dedifferentiating plant cells enter cell division.
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Abbreviations
- AF:
-
actin filament
- BDM:
-
2,3-butanedione monoxime
- GFP:
-
green-fluorescent protein
- IVS:
-
internal vacuolar structure
- LatB:
-
latrunculin B
- MT:
-
microtubule
- NEM:
-
N-ethylmaleimide
- Ory:
-
oryzalin
- TIP:
-
tonoplast-intrinsic protein
- TVS:
-
transvacuolar strands
References
PC Bethke RL Jones (2000) ArticleTitleVacuoles and prevacuolar compartments Curr Opin Plant Biol 3 469–475 Occurrence Handle11074377 Occurrence Handle10.1016/S1369-5266(00)00115-1 Occurrence Handle1:STN:280:DC%2BD3M%2FmtFaqtA%3D%3D
M Chalfie Y Tu G Euskirchen WW Ward DC Prasher (1994) ArticleTitleGreen fluorescent protein as a marker for gene expression Science 263 802–805 Occurrence Handle8303295 Occurrence Handle10.1126/science.8303295 Occurrence Handle1:CAS:528:DyaK2cXitFWkurc%3D
PFL Chang B Damsz AK Kononowicz M Reuveni Z Chen Y Xu K Hedges CC Tseng NK Singh ML Binzel ML Narasimhan PM Hasegawa RA Bressan (1996) ArticleTitleAlterations in cell membrane structure and expression of a membrane-associated protein after adaptation to osmotic stress Physiol Plant 98 505–516 Occurrence Handle10.1111/j.1399-3054.1996.tb05705.x Occurrence Handle1:CAS:528:DyaK28XntFKqt7w%3D
SR Cutler DW Ehrhardt JS Griffitts CR Somerville (2000) ArticleTitleRandom GFP::cDNA fusions enable visualization of subcellular structures in cells of Arabidopsis at a high frequency Proc Natl Acad Sci USA 97 3718–3723 Occurrence Handle10737809 Occurrence Handle10.1073/pnas.97.7.3718 Occurrence Handle1:CAS:528:DC%2BD3cXitlajsr0%3D
G Ferreira P Artigas G Pizarro G Brum (1997) ArticleTitleButanedione monoxime promotes voltage-dependent inactivation of L-type calcium channels in heart. Effects on gating currents J Mol Cell Cardiol 29 777–787 Occurrence Handle9140834 Occurrence Handle10.1006/jmcc.1996.0321 Occurrence Handle1:CAS:528:DyaK2sXivVSgtbw%3D
AE Franklin WZ Cande (1999) ArticleTitleNuclear organization and chromosome segregation Plant Cell 11 523–534 Occurrence Handle10213775 Occurrence Handle10.1105/tpc.11.4.523 Occurrence Handle1:CAS:528:DyaK1MXjtFSrtbs%3D
F Grolig ES Pierson (2000) Cytoplasmic streaming: from flow to track C Staiger F Baluška P Volkmann P Barlow (Eds) Actin: a dynamic framework for multiple plant cell functions Kluwer Academic Dordrecht 165–190
GR Hicks E Rojo S Hong DG Carter NV Raikhel (2004) ArticleTitleGerminating pollen has tubular vacuoles, displays highly dynamic vacuole biogenesis, and requires VACUOLESS1 for proper function Plant Physiol 134 1227–1239 Occurrence Handle14988481 Occurrence Handle10.1104/pp.103.037382 Occurrence Handle1:CAS:528:DC%2BD2cXis1Sjtb0%3D
A Hoffmann A Nebenführ (2004) ArticleTitleDynamic rearrangements of transvacuolar strands in BY-2 cells imply a role of myosin in remodeling the plant actin cytoskeleton Protoplasma 224 201–210 Occurrence Handle15614481 Occurrence Handle10.1007/s00709-004-0068-0 Occurrence Handle1:CAS:528:DC%2BD2cXhtFWqsLfI
PJ Hussey M Yuan G Calder S Khan CW Lloyd (1998) ArticleTitleMicroinjection of pollen-specific actin-depolymerizing factor, ZmADF1, reorients F-actin strands in Tradescantia stamen hair cells Plant J 14 353–357 Occurrence Handle10.1046/j.1365-313X.1998.00122.x Occurrence Handle1:CAS:528:DyaK1cXjslOlsr8%3D
GY Jauh TE Phillips JC Rogers (1999) ArticleTitleTonoplast intrinsic protein isoforms as markers for vacuolar functions Plant Cell 11 1867–1882 Occurrence Handle10521518 Occurrence Handle10.1105/tpc.11.10.1867 Occurrence Handle1:CAS:528:DyaK1MXntFCgt7g%3D
KN Kao MR Michayluk (1975) ArticleTitleNutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media Planta 126 105–110 Occurrence Handle10.1007/BF00380613 Occurrence Handle1:CAS:528:DyaE28Xjs1Ol
J Katsuta Y Hashiguchi H Shibaoka (1990) ArticleTitleThe role of the cytoskeleton in the positioning of the nucleus in premitotic tobacco BY-2 cells J Cell Sci 95 413–422
N Kutsuna S Hasezawa (2002) ArticleTitleDynamic organization of vacuolar and microtubule structures during cell cycle progression in synchronized tobacco BY-2 cells Plant Cell Physiol 43 965–973 Occurrence Handle12354913 Occurrence Handle10.1093/pcp/pcf138 Occurrence Handle1:CAS:528:DC%2BD38XntlGjur8%3D
S Liebe D Menzel (1995) ArticleTitleActomyosin-based motility of endoplasmic reticulum and chloroplasts in Vallisneria mesophyll cells Biol Cell 85 207–222 Occurrence Handle8785522 Occurrence Handle10.1016/0248-4900(96)85282-8 Occurrence Handle1:CAS:528:DyaK28Xkt1Sru74%3D
S Liebe H Quader (1994) ArticleTitleMyosin in onion (Allium cepa) bulb scale epidermal cells: involvement in dynamics of organelles and endoplasmic reticulum Plant Physiol 90 114–124 Occurrence Handle10.1111/j.1399-3054.1994.tb02200.x Occurrence Handle1:CAS:528:DyaK2cXhvVOhsbc%3D
F Marty (1999) ArticleTitlePlant vacuoles Plant Cell 11 587–600 Occurrence Handle10213780 Occurrence Handle10.1105/tpc.11.4.587 Occurrence Handle1:CAS:528:DyaK1MXjtFSrur4%3D
TM Molchan AH Valster PK Hepler (2002) ArticleTitleActomyosin promotes cell plate alignment and late lateral expansion in Tradescantia stamen hair cells Planta 214 683–693 Occurrence Handle11882936 Occurrence Handle10.1007/s004250100672 Occurrence Handle1:CAS:528:DC%2BD38Xhs1yqs7g%3D
Y Moriyasu M Hattori GY Jauh JC Rogers (2003) ArticleTitleAlpha tonoplast intrinsic protein is specifically associated with vacuole membrane involved in an autophagic process Plant Cell Physiol 44 795–802 Occurrence Handle12941871 Occurrence Handle10.1093/pcp/pcg100 Occurrence Handle1:CAS:528:DC%2BD3sXmvVeisbg%3D
A Nebenführ LA Gallagher TG Dunahay JA Frohlick AM Mazurkiewicz JB Meehl LA Staehelin (1999) ArticleTitleStop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system Plant Physiol 121 1127–1142 Occurrence Handle10594100 Occurrence Handle10.1104/pp.121.4.1127
E Ono M Hatayama Y Isono T Sato R Watanabe K Yonekura-Sakakibara M Fukuchi-Mizutani Y Tanaka T Kusumi T Nishino T Nakayama (2006) ArticleTitleLocalization of a flavonoid biosynthetic polyphenol oxidase in vacuoles Plant J 45 133–143 Occurrence Handle16367960 Occurrence Handle10.1111/j.1365-313X.2005.02625.x Occurrence Handle1:CAS:528:DC%2BD28XhtV2it78%3D
KS Opalski H Schultheiss KH Kogel R Huckelhoven (2005) ArticleTitleThe receptor-like MLO protein and the RAC/ROP family G-protein RACB modulate actin reorganization in barley attacked by the biotrophic powdery mildew fungus Blumeria graminis f.sp. hordei Plant J 41 291–303 Occurrence Handle15634205 Occurrence Handle10.1111/j.1365-313X.2004.02292.x Occurrence Handle1:CAS:528:DC%2BD2MXhtVWit7k%3D
TP Pasternak E Prinsen F Ayaydin P Miskolczi G Potters H Asard HA Van Onckelen D Dudits A Feher (2002) ArticleTitleThe role of auxin, pH, and stress in the activation of embryogenic cell division in leaf protoplast-derived cells of alfalfa Plant Physiol 129 1807–1819 Occurrence Handle12177494 Occurrence Handle10.1104/pp.000810 Occurrence Handle1:CAS:528:DC%2BD38Xmtl2gtLw%3D
J Pokorna K Schwarzerova S Zelenkova J Petrasek I Janotova V Capkova Z Opatrny (2004) ArticleTitleSites of actin filament initiation and reorganization in cold-treated tobacco cells Plant Cell Environ 27 641–653 Occurrence Handle10.1111/j.1365-3040.2004.01186.x Occurrence Handle1:CAS:528:DC%2BD2cXkvF2lsL4%3D
D Reisen N Leborgne-Castel C Ozalp F Chaumont F Marty (2003) ArticleTitleExpression of a cauliflower tonoplast aquaporin tagged with GFP in tobacco suspension cells correlates with an increase in cell size Plant Mol Biol 52 387–400 Occurrence Handle12856944 Occurrence Handle10.1023/A:1023961332391 Occurrence Handle1:CAS:528:DC%2BD3sXktFSmurc%3D
D Reisen F Marty N Leborgne-Castel (2005) ArticleTitleNew insights into the tonoplast architecture of plant vacuoles and vacuolar dynamics during osmotic stress BMC Plant Biol 5 13 Occurrence Handle16080795 Occurrence Handle10.1186/1471-2229-5-13
TL Rose L Bonneau C Der D Marty-Mazars F Marty (2006) ArticleTitleStarvation-induced expression of autophagy-related genes in Arabidopsis Biol Cell 98 53–67 Occurrence Handle16354162 Occurrence Handle10.1042/BC20040516 Occurrence Handle1:CAS:528:DC%2BD28XhsFeis74%3D
C Saito MT Morita T Kato M Tasaka (2005) ArticleTitleAmyloplasts and vacuolar membrane dynamics in the living graviperceptive cell of the Arabidopsis inflorescence stem Plant Cell 17 548–558 Occurrence Handle15689424 Occurrence Handle10.1105/tpc.104.026138 Occurrence Handle1:CAS:528:DC%2BD2MXhsFKqs78%3D
M Saitoh T Ishikawa S Matsushima M Naka H Hidaka (1987) ArticleTitleSelective inhibition of catalytic activity of smooth muscle myosin light chain kinase J Biol Chem 262 7796–7801 Occurrence Handle3108259 Occurrence Handle1:CAS:528:DyaL2sXksFCksLg%3D
T Selga (2003) ArticleTitleChanges of the secretory system of leaf epidermal and mesophyll cells during stress and programmed cell death Acta Univ Latv 662 51–58
MB Sheahan RJ Rose DW McCurdy (2004a) ArticleTitleOrganelle inheritance in plant cell division: the actin cytoskeleton is required for unbiased inheritance of chloroplasts, mitochondria and endoplasmic reticulum in dividing protoplasts Plant J 37 379–390 Occurrence Handle10.1046/j.1365-313X.2003.01967.x Occurrence Handle1:CAS:528:DC%2BD2cXhvFejtr8%3D
MB Sheahan CJ Staiger RJ Rose DW McCurdy (2004b) ArticleTitleA 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 Occurrence Handle10.1104/pp.104.049411 Occurrence Handle1:CAS:528:DC%2BD2MXjtValsw%3D%3D
MB Sheahan DW McCurdy RJ Rose (2005) ArticleTitleMitochondria as a connected population: ensuring continuity of the mitochondrial genome during plant cell dedifferentiation through massive mitochondrial fusion Plant J 44 744–755 Occurrence Handle16297067 Occurrence Handle10.1111/j.1365-313X.2005.02561.x Occurrence Handle1:CAS:528:DC%2BD2MXhtlWltrnJ
T Shimmen M Hamatani S Saito E Yokota T Mimura N Fusetani H Karaki (1995) ArticleTitleRoles of actin filaments in cytoplasmic streaming and organization of transvacuolar strands in root hair cells of Hydrocharis Protoplasma 185 188–193 Occurrence Handle10.1007/BF01272859 Occurrence Handle1:CAS:528:DyaK2MXnvFSgsrY%3D
IM Tolić-Nørrelykke L Sacconi C Stringari I Raabe FS Pavone (2005) ArticleTitleNuclear and division-plane positioning revealed by optical micromanipulation Curr Biol 15 1212–1216 Occurrence Handle16005294 Occurrence Handle10.1016/j.cub.2005.05.052
M Tominaga E Yokota L Vidali S Sonobe PK Hepler T Shimmen (2000) ArticleTitleThe role of plant villin in the organization of the actin cytoskeleton, cytoplasmic streaming and the architecture of the transvacuolar strand in root hairs of Hydrocharis Planta 210 836–843 Occurrence Handle10805457 Occurrence Handle10.1007/s004250050687 Occurrence Handle1:CAS:528:DC%2BD3cXit1Gjtbg%3D
JA Traas JH Doonan DJ Rawlins PJ Shaw J Watts CW Lloyd (1987) ArticleTitleAn actin network is present in the cytoplasm throughout the cell cycle of carrot cells and associates with the dividing nucleus J Cell Biol 105 387–395 Occurrence Handle2440896 Occurrence Handle10.1083/jcb.105.1.387 Occurrence Handle1:CAS:528:DyaL2sXltlWiu70%3D
K van Gestel RH Köhler JP Verbelen (2002) ArticleTitlePlant mitochondria move on F-actin, but their positioning in the cortical cytoplasm depends on both F-actin and microtubules J Exp Bot 53 659–667 Occurrence Handle11886885 Occurrence Handle10.1093/jexbot/53.369.659 Occurrence Handle1:CAS:528:DC%2BD38XitlCks7w%3D
S-X Xu (1992) ArticleTitleConfocal microscopic observations on actin filament distribution in lily pollen protoplasts Acta Bot Sin 34 907–911 Occurrence Handle1:CAS:528:DyaK3sXkvFWhur8%3D
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Correspondence and reprints: School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia.
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Video1.avi
Video 1. Vacuolar architecture in mesophyll of intact A. thaliana seedlings. A 66 μm deep confocal section through immature leaf tissue showing limited internal vacuolar structure in mesophyll cells. Video runs at 6 μm/s, with section interval of 1 μm. Bar: 20 μm
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Video 2. Dynamic transvacuolar strands in epidermal cells of the hypocotyl. Images captured at 6 s intervals. Video runs at 30× actual speed. Bar: 20 μm
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Sheahan, M., Rose, R. & McCurdy, D. Actin-filament-dependent remodeling of the vacuole in cultured mesophyll protoplasts. Protoplasma 230, 141–152 (2007). https://doi.org/10.1007/s00709-006-0236-5
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DOI: https://doi.org/10.1007/s00709-006-0236-5