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The effects of microtubule and microfilament disrupting agents on cytoskeletal arrays and wall deposition in developing cotton fibers

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The effects of various cytoskeletal disrupting agents (cholchicine, oryzalin, trifluralin, taxol, cytochalasins B and D) on microtubules, microfilaments and wall microfibril deposition were monitored in developing cotton fibers, using immunocytochemical and fluorescence techniques. Treatment with 10−4 M colchicine, 10−6 M trifluralin or 10−6 M oryzalin resulted in a reduction in the number of microtubules, however, the “drug-stable” microtubules still appear to influence wall deposition. Treatment with 10−5 M taxol increased the numbers of microtubules present within 15 minutes of application. New microtubules were aligned parallel to the existing ones, however, some evidence of random arrays was observed. Microtubules stabilized with taxol appeared to function in wall organization but do not undergo normal re-orientations during development. Microtubule disrupting agent had no detectable affect on the microfilament population. Exposure to either 4×10−5 M cytochalasin B or 2×10−6M cytochalasin D resulted in a disruption of microfilaments and a re-organization of microtubule arrays. Treatment with either cytochalasin caused a premature shift in the orientation of microtubules in young fibers, whereas in older fibers the microtubule arrays became randomly organized. These observations indicate that microtubule populations during interphase are heterogeneous, differing at least in their susceptibility to disruption by depolymerizing agents. Changes in microtubule orientation (induced by cytochalasin) indicate that microfilaments may be involved in regulating microtubule orientation during development.

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References

  • Beasley CA, Ting IP (1973) The effects of plant growth substances on in vitro fiber development in fertilized cotton ovules. Amer J Bot 60: 130–139

    Google Scholar 

  • Bajer AS, Mole-Bajer J (1986) Drugs with colchicine-like effects that specifically disassemble plant but not animal microtubules. Ann NY Acad Sci 466: 767–784

    Google Scholar 

  • Cyr RJ, Palevitz BA (1989) Microtubule-binding proteins from carrot. I. Initial characterization and microtubule bundling. Planta 177: 245–260

    Google Scholar 

  • —, Bustos MM, Guiltinan MJ, Fosket DE (1987) Developmental modulation of tubulin protein and mRNA levels during somatic embryogenesis in cultured carrot cells. Planta 171: 365–376

    Google Scholar 

  • Dawson PJ, Lloyd CW (1985) Identification of multiple tubulins in taxol microtubules purified from carrot suspension cells. EMBO J 4: 2451–45

    Google Scholar 

  • Derksen J, Traas JA (1984) Growth of tobacco pollen in vitro: effects of drugs interfering with the cytoskeleton. In: Willemse MTM, Went JL (eds) Proceedings 8th International Symposium Sexual Reproduction in Seed Plants, Ferns, and Mosses. Pudoc, Wageningen, pp 64–67

  • — — Oosterdorp T (1986) Distribution of actin filaments in differentiating cells ofEquisetum hyemale root tips. Plant Sci 43: 77–81

    Google Scholar 

  • Falconer MM, Seagull RW (1985 a) Immunofluorescent and calcofluor white staining of developing tracheary elements inZinnia elegans L. suspension cultures. Protoplasma 125: 190–198

    Google Scholar 

  • — — (1985 b) Xylogenesis in tissue culture: taxol effects on microtubule reorientation and lateral association in differentiating cells. Protoplasma 128: 157–166

    Google Scholar 

  • — — (1987) Amiprophos-methyl (APM): a rapid, reversible, antimicrotubule agent for plant cell cultures. Protoplasma 136: 118–124

    Google Scholar 

  • Farrell KW, Wilson L (1980) Proposed mechanism for colchicine poisoning of microtubules reassembled in vitro fromStongylocintrotus purpatus sperm tail outer doublet tubulin. Biochemistry 9: 3466–3472

    Google Scholar 

  • Fukuda H, Kobayashi H (1989) Dynamic organization of the cytoskeleton during tracheary-element differentiation. Dev Growth Different 31: 9–16

    Google Scholar 

  • Galatis B (1977) Differentiation of stomatal meristemoids and guard cell mother cells into guard-like cells inVigna sinensis leaves after colchicine treatment. Planta 136: 103–114

    Google Scholar 

  • Gunning BES, Hardham AR (1982) Microtubules. Annu Rev Plant Physiol 33: 651–698

    Google Scholar 

  • Hardham AR, McCully ME (1982) Preprogramming of cells following wounding in pea (Pisum sativum L.) root. II. The effects of caffeine and colchicine on the development of new vascular elements. Protoplasma 112: 152–166

    Google Scholar 

  • —, Gunning BES (1979) Interpolation of microtubules into cortical arrays during cell elongation and differentiation in roots ofAzolla pinnata. J Cell Sci 37: 411–442

    Google Scholar 

  • Hartwig JH, Stossel TP (1979) Cytochalasin B and the structure of actin gels. J Mol Biol 134: 539–553

    Google Scholar 

  • Heath IB, Seagull RW (1982) Oriented cellulose microfibrils and the cytoskeleton: a critical comparison of models. In: Lloyd CW (ed) The cytoskeleton in plant growth and development. Academic Press, London, pp 162–182

    Google Scholar 

  • Hensel W (1986) Cytodifferentiation in polar plant cells. Use of antimicro tubular agents during the differentiation of statocytes from cress roots (Lepidium sativum L.). Planta 169: 293–303

    Google Scholar 

  • Hepler PK, Palevitz BA (1974) Microtubules and microfilaments. Annu Rev Plant Physiol 25: 309–362

    Google Scholar 

  • Hess FD, Bayer DE (1977) Binding of the herbicide trifluralin toChlamydomonas flagellar tubulin. J Cell Sci 24: 351–360

    Google Scholar 

  • Hussey PJ, Gull K (1985) Multiple isotypes of A and B tubulin in the plantPhaseolus vulgaris. FEBS Lett 181: 113–118

    Google Scholar 

  • —, Traas JA, Gull K, Lloyd CW (1987) Isolation of cytoskeletons from synchronized plant cells: the interphase microtubule array utilizes multiple tubulin isotypes. J Cell Sci 88: 225–230

    Google Scholar 

  • Juniper BE, Lawton JR (1979) The effect of caffeine, different fixation regimes and low temperature on microtubules in the cells of higher plants. Planta 145: 411–416

    Google Scholar 

  • Kakimoto T, Shibaoka H (1987) Actin filaments and microtubules in the pre-prophase band and phragmoplast of tobacco cells. Protoplasma 140: 151–156

    Google Scholar 

  • Katsuta J, Shibaoka H (1988) The roles of the cytoskeleton and the cell wall in nuclear positioning in tobacco BY-2 cells. Plant Cell Physiol 29: 403–413

    Google Scholar 

  • Kloth R (1989) Changes in the level of tubulin subunits during development of cotton (Gossypium hirsutum) fiber. Physiol Plant 76: 37–41

    Google Scholar 

  • Kobayashi H, Fukuda H, Shibaoka H (1989) Interrelation between the spatial disposition of actin filaments and microtubules during the differentiation of tracheary elements in culturedZinnia elegans. Protoplasma 143: 29–37

    Google Scholar 

  • Lancelle SA, Hepler PK (1987) Ultrastructure of the cytoskeleton in freeze-substituted pollen tubes ofNicotiana alata. Protoplasma 140: 141–150

    Google Scholar 

  • Lloyd CW (1984) Toward a dynamic helical model for the influence of microtubules on wall patterns in plants. Int Rev Cytol 86: 1–51

    Google Scholar 

  • MacLean-Fletcher S, Pollard TD (1980) Mechanism of action of cytochalasin B on actin. Cell 20: 329–341

    Google Scholar 

  • Margolis RL, Wilson L (1977) Addition of colchicine-tubulin complex to microtubule ends: the mechanism of substoichiometric colchicine poisoning. Proc Natl Acad Sci USA 74: 3466–3470

    Google Scholar 

  • Meyer Y, Herth W (1978) Chemical inhibition of cell wall formation and cytokinesis, but not nuclear division, in protoplasts inNicotiana tabacum L. cultivated in vitro. Planta 142: 253–262

    Google Scholar 

  • Morejohn LC, Fosket DE (1984) Inhibition of plant microtubule polymerization in vitro by the phosphoric amide herbicide Amiprophos-methyl. Science 224: 874–876

    Google Scholar 

  • —, Bureau TE, Tocchi LP, Fosket DE (1984) Tubulins from different higher plant species are immunologically nonidentical and bind colchicine differentially. Proc Natl Acad Sci USA 81: 1440–1444

    Google Scholar 

  • — —, Mole-Bajer J, Bajer AS, Fosket DE (1987) Oryzalin, a dinitroaniline herbicide, binds to plant tubulin and inhibits microtubule polymerization in vitro. Planta 172: 252–264

    Google Scholar 

  • Nelms BJ, Preston RD, Ashworth D (1973) A possible function of microtubules suggested by their distribution in rubbery wood. J Cell Sci 37: 411–424

    Google Scholar 

  • Palevitz BA (1980) Comparative effects of phalloidin and cytochalasin B on motility and morphogenesis inAllium. Can J Bot 58: 772–785

    Google Scholar 

  • — (1987 a) Actin in the preprophase band ofAllium cepa. J Cell Biol 104: 1515–1519

    Google Scholar 

  • — (1987 b) Accumulation of F-actin during cytokinesis inAllium. Correlation with microtubule disruption and the effects of drugs. Protoplasma 141: 24–32

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Pickett-Heaps JD (1967) The effects of colchicine on the ultrastructure of dividing plant cells, xylem wall differentiation and distribution of cytoplasmic microtubules. Dev Biol 15: 206–236

    Google Scholar 

  • Pierson ES (1988) Rhodamine-phalloidin staining of F-actin in pollen after dimethylsulfoxide permeabilization. Sex Plant Reprod 1: 83–87

    Google Scholar 

  • Quader H, Filner P (1980) The action of antimitotic herbicides on flagellar regeneration inChlamydomonas reinhardtii: a comparison with the action of colchicine. Eur J Cell Biol 21: 301–304

    Google Scholar 

  • Ridge R (1988) Freeze-substitution improves the ultrastructural preservation of legume root hairs. Bot Mag Tokyo 101: 427–441

    Google Scholar 

  • Schiff PB, Fant J Horowitz SB (1979) Promotion of microtubule assembly in vitro by taxol. Nature 277: 665–667

    Google Scholar 

  • Schiff PB, Horowitz SB (1980) Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci USA 77: 1561–1565

    Google Scholar 

  • Schmit A-C, Lambert A-M (1988) Plant actin filament and microtubule interactions during anaphase-telophase transition: effects of antagonist drugs. Biol Cell 64: 309–319

    Google Scholar 

  • Seagull RW (1986) Changes in microtubule organization and microfibril orientation during in vitro cotton fiber development: an immunofluorescent study. Can J Bot 64: 1373–1381

    Google Scholar 

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

    Google Scholar 

  • - (1989 b) Changes in microtubule arrays during cotton fiber development. In: Bailey GW (ed) Proceedings of the 47th Annual Meeting of the Electron Microscopy Society of America, San Antonio, Texas, pp 760–761

  • — (1990) The role of the cytoskeleton in cellulose synthesis. In: Haigler C, Weimer PO (eds) Biogenesis and biodégradation of cellulose and cellulosic materials. Marcel Dekker, New York (in press)

    Google Scholar 

  • —, Heath IB (1980 a) The differential effects of cytochalasin B on microfilament populations and cytoplasmic streaming. Protoplasma 103: 231–240

    Google Scholar 

  • — (1980b) The organization of cortical microtubule arrays in the radish root hair. Protoplasma 103: 205–229

    Google Scholar 

  • —, Falconer MM, Weerdenberg CA (1987) Microfilaments: dynamic arrays in higher plant cells. J Cell Biol 104: 995–1004

    Google Scholar 

  • —, Holland S, Gibson DM (1989) Isolation of cytoskeletal proteins from cotton (Gossypium hirsutum) suspension culture cells. J Cell Biol 109: 78 a

    Google Scholar 

  • Sheterline P (1983) Mechanisms of cell motility: molecular aspects of contractility. Academic Press, London, pp 167–175

    Google Scholar 

  • Simmonds DH, Seagull RW, Setterfield G (1985) Evaluation of techniques for immunofluorescent staining of microtubules in cultured plant cells. J Histochem Cytochem 33: 345–352

    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

    Google Scholar 

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

    Google Scholar 

  • Tiwari CC, Gunning BES (1986) Colchicine inhibits plasmodium formation and disrupts pathway of sporopollenin secretion in the anther tapetum ofTradescantia virginiana L. Protoplasma 114: 115–128

    Google Scholar 

  • —, Polito VS (1988) Organization of the cytoskeleton in pollen tubes ofPyrus communis: a study employing conventional and freezesubstitution electron microscopy, immunofluorescence and rhodamine-phalloidin. Protoplasma 147: 100–112

    Google Scholar 

  • Traas JA, Doonan JH, Rawlins 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

    Google Scholar 

  • Vaughn MA, Vaughn KC (1987) Effects of microfilament disrupters on microfilament distribution and morphology in maize root cells. Histochemistry 87: 129–137

    Google Scholar 

  • Volkmann D, Czaja AWP (1981) Reversible inhibition of secretion in root cap cells of cress after treatment with cytochalasin B. Exp Cell Res 135: 229–236

    Google Scholar 

  • Waterkyn L (1985) Light microscopy of the cotton fiber. Tech Monogr Belgian Cotton Res Council. International Institute of Cotton, Manchester, UK

    Google Scholar 

  • Weerdenberg CA, Falconer MM, Setterfield G, Seagull RW (1986) Effects of taxol on microtubule arrays in cultured higher plant cells. Cell Motil Cytoskeleton 6: 469–478

    Google Scholar 

  • Yatsu LY, Jacks TJ (1981) An ultrastructural study of the relationship between microtubules and microfibrils in cotton (Gossypium hirsutum L.) cell wall reversals. Amer J Bot 68: 771–777

    Google Scholar 

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Seagull, R.W. The effects of microtubule and microfilament disrupting agents on cytoskeletal arrays and wall deposition in developing cotton fibers. Protoplasma 159, 44–59 (1990). https://doi.org/10.1007/BF01326634

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