Skip to main content
SpringerLink
Log in

Cortical actin filaments fragment and aggregate to form chloroplast-associated and free F-actin rings in mechanically isolatedZinnia mesophyll cells

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

Changes in F-actin organization following mechanical isolation ofZinnia mesophyll cells were documented by rhodamine-phalloidin staining. Immediately after isolation, most cells contained irregular cortical actin fragments of varying lengths, and less than 5% of cells contained intact cortical filaments. During the first 8 h of culture, filament fragments were replaced by actin rings, stellate actin aggregates, and bundled filament fragments. Some of these aggregates had no association with organelles (“free” actin aggregates). Other aggregates were associated with chloroplasts, which changed in shape and location at the same time actin aggregates appeared. F-actin was concentrated within or around the nucleus in a small percentage of cells. After 12 h in culture, the percentage of cells with free actin rings and chloroplast-associated actin aggregates began to decline and the percentage of cells having intact cortical actin filaments increased greatly. Intermediate images were recorded that strongly indicate that free actin rings, chloroplast-associated actin rings, and other actin aggregates self-assemble by successive bundling of actin filament fragments. The fragmentation and bundling of F-actin observed in mechanically isolatedZinnia cells resembles changes in F-actin distribution reported after diverse forms of cell disturbance and appears to be an example of a generalized response of the actin cytoskeleton to cell stress.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

FITC:

fluorescein isothiocyanate

MBS:

m-maleimidobenzoic acid N-hydroxysuccinimide ester

RhPh:

tetramethylrhodamine isothiocyanate-phalloidin

References

  • Alfano F, Russell A, Gambardella R, Duckett JG (1993a) The actin cytoskeleton of the liverwortRiccia fluitans: effects of cytochalasin B and aluminium ions on rhizoid tip growth. J Plant Physiol 142: 569–574

    Google Scholar 

  • —, Duckett JG, Gambardella R (1993b) Circular and spiral F-actin associated with chloroplasts in liverwort thalli and in gametophyte cells ofEquisetum andPteridium. G Bot Ital 127: 292–294

    Google Scholar 

  • Allen NS, Kubinski DJ (1995) Actin dynamics and a putative gelsolin inMedicago sativa (alfalfa). Mol Biol Cell Suppl 6: 22A

    Google Scholar 

  • Borovikov YS, Norman JC, Price LS, Weeds A, Koffer A (1995) Secretion from permeabilized mast cells is enhanced by addition of gelsolin: contrasting effects of endogenous gelsolin. J Cell Sci 108: 657–666

    PubMed  Google Scholar 

  • Cande WZ, Goldsmith MHM, Ray PM (1973) Polar auxin transport and auxin-induced elongation in the absence of cytoplasmic streaming. Planta 111: 279–296

    Google Scholar 

  • Cho S-O, Wick SM (1990) Distribution and function of actin in the developing stomatal complex of winter rye (Secale cereale cv. Puma). Protoplasma 157: 154–164

    Google Scholar 

  • Collings DA, Wasteneys GO, Williamson RE (1995) Cytochalasin rearranges cortical actin of the algaNitella into short, stable rods. Plant Cell Physiol 36: 765–772

    Google Scholar 

  • — —, Miyazaki M, Williamson RE (1994) Elongation factor 1α is a component of the subcortical actin bundles of characean algae. Cell Biol Int Rep 18: 1019–1024

    Google Scholar 

  • Cooper JA (1987) Effects of cytochalasin and phalloidin on actin. J Cell Biol 105: 1473–1478

    PubMed  Google Scholar 

  • —, Bryan J, Schwab B III, Frieden C, Loftus DJ, Elson EL (1987) Microinjection of gelsolin into living cells. J Cell Biol 104: 491–501

    PubMed  Google Scholar 

  • Dong X-J, Ryu J-H, Takagi S, Nagai R (1996) Dynamic changes in the organization of microfilaments associated with the photocontrolled motility of chloroplasts in epidermal cells ofVallisneria. Protoplasma 195 (in press)

  • Fukuda H, Komamine A (1980) Establishment of an experimental system for the study of tracheary element differentiation from single cells isolated from the mesophyll ofZinnia elegans. Plant Physiol 65: 57–60

    Google Scholar 

  • Fukui Y, Katsumaru H (1980) Dynamics of nuclear actin bundle induction by dimethyl sulfoxide and factors affecting its development. J Cell Biol 84: 131–140

    PubMed  Google Scholar 

  • Garbary DJ, McDonald AR (1996) Fluorescent labelling of the cytoskeleton inCeramium strictum (Rhodophyta). J Phycol 32: 85–93

    Google Scholar 

  • Goddard RH, La Claire JW II (1991a) Calmodulin and wound healing in the coenocytic green algaErnodesmis verticillata (Kutzing) Borgesen: immunofluorescence and effects of antagonists. Planta 183: 281–293

    Google Scholar 

  • — — (1991b) Calmodulin and wound healing in the coenocytic green algaErnodesmis verticillata (Kutzing) Borgesen: ultrastructure of the cortical cytoskeleton and immunogold labeling. Planta 186: 17–26

    Google Scholar 

  • Goodbody KC, Lloyd CW (1990) Actin filaments line up acrossTradescantia epidermal cells, anticipating wound-induced division planes. Protoplasma 157: 92–101

    Google Scholar 

  • Grabski S, Xie XG, Holland JF, Schindler M (1994) Lipids trigger changes in the elasticity of the cytoskeleton in plant cells: a cell optical displacement assay for live cell measurements. J Cell Biol 126: 713–726

    PubMed  Google Scholar 

  • Green PB (1994) Connecting gene and hormone action to form, pattern and organogenesis: biophysical transductions. J Exp Bot 45: 1775–1788

    Google Scholar 

  • Gross P, Julius C, Schmelzer E, Hahlbrock K (1993) Translocation of cytoplasm and nucleus to fungal penetration sites is associated with depolymerization of microtubules and defence gene activation in infected, cultured parsley cells. EMBO J 12: 1735–1744

    PubMed  Google Scholar 

  • Harold RL, Harold FM (1992) Configuration of actin microfilaments during sporangium development inAchlya bisexualis: comparison of two staining protocols. Protoplasma 171: 110–116

    Google Scholar 

  • Hasek J, Streiblova E (1994) Flunarizine affects F-actin patterns inMucor rouxii germlings. Can J Microbiol 40: 730–735

    Google Scholar 

  • Hasezawa S, Nagata T, Syono K (1988) The presence of ring formed actin filaments in plant cells. Protoplasma 146: 61–63

    Google Scholar 

  • He Y, Wetzstein HY (1995) Fixation induces differential tip morphology and immunolocalization of the cytoskeleton in pollen tubes. Physiol Plant 93: 757–763

    Google Scholar 

  • Higashi-Fujime S (1980) Active movement in vitro of bundles of microfilaments isolated fromNitella cell. J Cell Biol 87: 569–578

    PubMed  Google Scholar 

  • Huckriede A, Fuchtbauer A, Hinssen H, Chaponnier C, Weeds A, Jockusch BM (1990) Differential effects of gelsolins on tissue culture cells. Cell Motil Cytoskeleton 16: 229–238

    PubMed  Google Scholar 

  • Hush JM, Overall RL (1992) Re-orientation of cortical F-actin is not necessary for wound-induced microtubule re-orientation and cell polarity establishment. Protoplasma 169: 97–106

    Google Scholar 

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

    Google Scholar 

  • — — (1992b) Re-organization of the cortical cytoskeleton in tip-growing fern protonemal cells during phytochrome-mediated phototropism and blue light-induced apical swelling. Protoplasma 166: 35–41

    Google Scholar 

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

    Google Scholar 

  • Kaminskyj SGW, Heath IB (1994) A comparison of techniques for localizing actin and tubulin in hyphae ofSaprolegnia ferax. J Histochem Cytochem 42: 523–530

    PubMed  Google Scholar 

  • Knight BW, Roberts AW (1994) Palisade mesophyll cell expansion during leaf development inZinnia elegans (Asteraceae). Am J Bot 81: 609–615

    Google Scholar 

  • Kobayashi H, Fukuda H, Shibaoka H (1988) Interrelationship between the spatial disposition of actin filaments and microtubules during the differentiation of tracheary elements in culturedZinnia cells. Protoplasma 143: 29–37

    Google Scholar 

  • Kobayashi I, Kobayashi Y, Hardham AR (1994) Dynamic re-organization of microtubules and microfilaments in flax cells during the resistance response to flax rust infection. Planta 195: 237–247

    Google Scholar 

  • — —, Yamaoka N, Kunoh H (1992) Recognition of a pathogen and a nonpathogen by barley coleoptile cells. III. Responses of microtubules and actin filaments in barley coleoptile cells to penetration attempts. Can J Bot 70: 1815–1823

    Google Scholar 

  • Kohno T, Shimmen T (1987) Ca2+-induced fragmentation of actin filaments in pollen tubes. Protoplasma 141: 177–179

    Google Scholar 

  • Kuroiwa T, Suzuki K, Itou R, Toda K, O'Keefe TC, Kuroiwa H (1995) Mitochondria-dividing ring: ultrastructural basis for the mechanism of mitochondrial division inCyanidioschyzon merolae. Protoplasma 186: 12–23

    Google Scholar 

  • La Claire JW II (1989) Actin cytoskeleton in intact and wounded coenocytic green algae. Planta 177: 47–57

    Google Scholar 

  • Lancelle SA, Hepler PK (1991) Association of actin with cortical microtubules revealed by immunogold localization inNicotiana pollen tubes. Protoplasma 165: 167–172

    Google Scholar 

  • — — (1988) Cytochalasin-induced ultrastructural alterations inNicotiana pollen tubes. Protoplasma Suppl 2: 65–75

    Google Scholar 

  • Lehrer SS (1981) Damage to actin filaments by glutaraldehyde: protection by tropomyosin. J Cell Biol 90: 459–466

    PubMed  Google Scholar 

  • Liebe S, Quader H (1994) Myosin in onion (Allium cepa) bulb scale epidermal cells: involvement in dynamics of organelles and endoplasmic reticulum. Physiol Plant 90: 114–124

    Google Scholar 

  • McCurdy DW, Gunning BES (1990) Re-organization of cortical actin microfilaments and microtubules at preprophase and mitosis in wheat root-tip cells: a double label immunofluorescence study. Cell Motil Cytoskeleton 15: 76–87

    Google Scholar 

  • Masuda Y, Takagi S, Nagai R (1991) Protease-sensitive anchoring of microfilament bundles provides tracks for cytoplasmic streaming inVallisneria. Protoplasma 162: 151–159

    Google Scholar 

  • Menzel D (1994) Cell differentiation and the cytoskeleton inAcetabularia. New Phytol 128: 369–393

    Google Scholar 

  • — (1987) The cytoskeleton of the giant coenocytic green algaCaulerpa visualized by immunocytochemistry. Protoplasma 139: 71–76

    Google Scholar 

  • —, Schliwa M (1986) Motility in the siphonous green algaBryopsis. II. Chloroplast movement requires organized arrays of both microtubules and actin filaments. Eur J Cell Biol 40: 286–295

    PubMed  Google Scholar 

  • Meske V, Hartmann E (1995) Reorganization of microfilaments in protonemal tip cells of the mossCeratodon purpureus during the phototropic response. Protoplasma 188: 59–69

    PubMed  Google Scholar 

  • Miller DD, Scordilis SP, Hepler PK (1995) Identification and localization of three classes of myosins in pollen tubes ofLilium longiflorum andNicotiana alata. J Cell Sci 108: 2549–2563

    PubMed  Google Scholar 

  • Northcote DH (1995) Aspects of vascular tissue differentiation in plants: parameters that may be used to monitor the process. Int J Plant Sci 156: 245–256

    Google Scholar 

  • Ogawa S, Ueda K, Noguchi T (1995) Division apparatus of the chloroplast inNannochloris bacillaris (Chlorophyta). J Phycol 31: 132–137

    Google Scholar 

  • Palevitz BA (1988) Cytochalasin-induced reorganization of actin inAllium root cells. Cell Motil Cytoskeleton 9: 283–298

    Google Scholar 

  • Parthasarathy MV (1985) F-actin architecture in coleoptile epidermal cells. Eur J Cell Biol 39: 1–12

    Google Scholar 

  • —, Perdue TD, Witztum A, Alvernaz J (1985) Actin network as a normal component of the cytoskeleton in many vascular plant cells. Am J Bot 72: 1318–1323

    Google Scholar 

  • Peat L, Oliveira L (1994) Organization of the cytoskeleton in the coenocytic algaVaucheria longicaulis var.macounii: an experimental study. Protoplasma 177: 95–107

    Google Scholar 

  • Pierson ES, Derksen J, Traas JA (1986) Organization of microfilaments and microtubules in pollen tubes grown in vitro or in vivo in various angiosperms. Eur J Cell Biol 41: 14–18

    Google Scholar 

  • Pollard TD, Cooper JA (1986) Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem 55: 987–1035

    PubMed  Google Scholar 

  • Putnam-Evans C, Harmon AC, Palevitz BA, Fechheimer M, Cormier MJ (1989) Calcium-dependent protein kinase is localized with F-actin in plant cells. Cell Motil Cytoskeleton 12: 12–22

    Google Scholar 

  • Quader H (1990) Formation and disintegration of cisternae of the endoplasmic reticulum visualized in live cells by conventional fluorescence and confocal laser scanning microscopy: evidence for the involvement of calcium and the cytoskeleton. Protoplasma 155: 166–175

    Google Scholar 

  • —, Schnepf E (1989) Actin filament array during side branch initiation in protonema cells of the mossFunaria hygrometrica: an actin organizing center at the plasma membrane. Protoplasma 151: 167–170

    Google Scholar 

  • —, Hofmann A, Schnepf E (1989) Reorganization of the endoplasmic reticulum in epidermal cells of onion bulb scales after cold stress: involvement of cytoskeletal elements. Planta 177: 273–280

    Google Scholar 

  • Roberts AW, Haigler CH (1989) Rise in chlorotetracycline fluorescence accompanies tracheary element differentiation in suspension cultures ofZinnia. Protoplasma 152: 37–45

    Google Scholar 

  • Ryu J-H, Takagi S, Nagai R (1995) Stationary organization of the actin cytoskeleton inVallisneria: the role of stable microfilaments at the end walls. J Cell Sci 108: 1531–1539

    PubMed  Google Scholar 

  • Sameshima M, Chijiiwa Y, Kishi Y, Hashimoto Y (1994) Novel actin rods appeared in spores ofDictyostelium discoideum. Cell Struct Funct 19: 189–194

    PubMed  Google Scholar 

  • Seagull RW (1990) The effects of microtubule and microfilament disrupting agents on cytoskeletal arrays and wall deposition in developing cotton fibers. Protoplasma 159: 44–59

    Google Scholar 

  • — (1989) The plant cytoskeleton. Crit Rev Plant Sci 8: 131–167

    Google Scholar 

  • Sonobe S, Shibaoka H (1989) Cortical fine actin filaments in higher plant cells visualized by rhodamine-phalloidin after pretreatment withm-maleimidobenzoyl N-hydroxysuccinimide ester. Protoplasma 148: 80–86

    Google Scholar 

  • Tang X, Lancelle SA, Hepler PK (1989) Fluorescence microscopic localization of actin in pollen tubes: comparison of actin antibody and phalloidin staining. Cell Motil Cytoskeleton 12: 216–224

    PubMed  Google Scholar 

  • Tewinkel M, Kruse S, Quader H, Volkmann D, Sievers A (1989) Visualization of actin filament pattern in plant cells without prefixation. A comparison of differently modified phallotoxins. Protoplasma 149: 178–182

    Google Scholar 

  • Thimann KV, Reese K, Nachmias VT (1992) Actin and the elongation of plant cells. Protoplasma 171: 153–166

    Google Scholar 

  • Tiwari SC, Polito VS (1988) Spatial and temporal organization of actin during hydration, activation, and germination of pollen inPyrus communis L.: a population study. Protoplasma 147: 5–15

    Google Scholar 

  • Traas JA, Doonan JH, Rawlings DJ, Shaw PJ, Watts J, Lloyd CW (1987) An 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

    PubMed  Google Scholar 

  • Uhnak KS, Roberts AW (1995) Microtubule rearrangements accompanying dedifferentiation in mesophyll cultures ofZinnia elegans L. Protoplasma 189: 81–87

    Google Scholar 

  • Wasteneys GO, Williamson RE (1991) Endoplasmic microtubules and nucleus-associated actin rings inNitella internodal cells. Protoplasma 162: 86–98

    Google Scholar 

  • —, Collings DA, Gunning BES, Hepler PK, Menzel D (1996) Actin in living and fixed characean internodal cells: identification of a cortical array of fine actin strands and chloroplast actin rings. Protoplasma 190: 25–38

    Google Scholar 

  • Williamson RE, Hurley UA (1986) Growth and regrowth of actin bundles inChara: bundle assembly by mechanisms differing in sensitivity to cytochalasin. J Cell Sci 85: 21–32

    PubMed  Google Scholar 

  • Yahara I, Harada F, Sekita S, Yoshihira K, Natori S (1982) Correlation between effects of 24 different cytochalasins on cellular structures and cellular events and those on actin in vitro. J Cell Biol 92: 69–78

    PubMed  Google Scholar 

  • Yang W, Burkhart W, Cavallius J, Merrick WC, Boss WF (1993) Purification and characterization of a phosphatidylinositol 4-kinase activator in carrot cells. J Biol Chem 268: 392–398

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biological Sciences, Ranger Hall, University of Rhode Island, 02881-0816, Kingston, RI, USA

    Alexander O. Frost & Alison W. Roberts

Authors
  1. Alexander O. Frost
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alison W. Roberts
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Frost, A.O., Roberts, A.W. Cortical actin filaments fragment and aggregate to form chloroplast-associated and free F-actin rings in mechanically isolatedZinnia mesophyll cells. Protoplasma 194, 195–207 (1996). https://doi.org/10.1007/BF01882027

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01882027

Keywords

Search

Navigation