Abstract
Translocations of chloroplasts induced by blue light were investigated in both leaves and protoplasts isolated from leaf mesophyll of Nicotiana tabacum. In the leaf tissue, the responses of chloroplasts were similar to those observed in other, higher and lower plant species. Weak and strong light induced movements of chloroplasts towards cell walls perpendicular and parallel to the light direction, respectively. Treatment with cytochalasin D, an actin-disturbing agent, blocked the movements. This shows that actin is involved in the motile system of chloroplast translocation in tobacco. By monitoring the response of chloroplasts to light in isolated protoplasts, we addressed the question whether the presence of the cell wall is necessary for the translocations of chloroplasts to occur. In control protoplasts (isolated at room temperature from unstressed leaves), no clear light intensity-dependent changes were observed in chloroplast distribution pattern. In contrast, in protoplasts obtained from plants treated with 4 °C for 8 h the chloroplasts maintained their responsiveness to light. Atomic Force Microscopy was used to measure elastic properties of the protoplasts. Young’s modulus, which reflects rigidity of the material, was 10 times higher for protoplasts of the coldstressed plants as compared to those isolated from the control plants. The rigidity of protoplasts isolated from the plants treated with low temperature was reduced four-fold by exposure to cytochalasin D. It appears that the status of protoplast actin is a factor responsible for elasticity of protoplasts. We speculate that unknown, cold stress-induced factors, maintain the orientational movements due to anchorage of the actin cytoskeleton in the plasma membrane despite the cell wall removal.
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References
Akashi T., Kawasaki S. & Shibaoka H. 1990. Stabilization of cortical microtubules by the cell wall in cultured tobacco cell. Planta 182, 363–369.
Augustynowicz J. & Gabryś H. 1999. Chloroplast movement in fern leaves: correlation of movement dynamics and environmental flexibility of the species. Plant Cell & Environ 22, 1239–1248.
Canut H., Carrasco A., Galaud J-P., Cassan C., Bouyssou H., Vita N., Ferrara P. & Pont-Lezica R. 1998. High affinity RGD-binding sites at the plasma membrane of Arabidopsis thaliana links the cell wall. Plant J 16, 63–71.
Collings D.A., Wasteneys G.O. & Williamson RE. 1996. Actin-microtubule interactions in the alga Nitella: analysis of the mechanism by which microtubule depolymerization potentates cytochalasin’s effects on streaming. Protoplasma 191, 178–190.
Collings D.A., Asada T., Allen N.S. & Shibaoka H. 1998. Plasma membrane-associated actin in Bright Yellow 2 tobacco cells. Plant Physiol 118, 917–928.
Dong X.-J., Ryu J.-H. & Nagai R. 1996. Dynamic changes in the organisation of microfilaments associated with the photocontrolled motility of chloroplasts in epidermal cells of Vallisneria. Protoplasma 195, 18–24.
Edelman G.M. 1993. A golden age for adhesion. Cell Adhes Commun 1, 1–7.
Faik A., Laboure A.M., Gulino D., Mandaron P. & Falconet D. 1998. A plant surface protein sharing structural properties with animal integrins. Eur J Biochem 253, 552–559.
Fowler J.E. & Quatrano R.S. 1997. Plant cell morphogenesis: plasma membrane interactions with the cytoskeleton and cell wall. Annu Rev of Cell Dev Biol 13, 697–743.
Griffith L.M. & Pollard T.D. 1982. The interaction of actin filaments with microtubules and microtubule associated proteins. J Biol Chem 257, 9143–9151.
Guy C.L. 1990. Cold acclimation and freezing stress tolerance: role of protein metabolism. Annu Rev Plant Physiol 41, 187–223.
Haupt W. & Scheuerlein R. 1990. Chloroplast movement. Plant Cell & Environ 13, 596–614.
Haupt W. 1998. Chloroplast movement: from phenomenology to molecular biology. In: Progress in Botany, Vol 60 (eds K. Esser, J.W. Kadereit, U. Lüttge & M. Runge), pp. 3–36. Springer-Verlag, Berlin, Heidelberg, New York.
Ingber D.E. 1991. Integrins as mechanochemical transducers. Curr Opin Cell Biol 3, 841–848.
Ishitani M., Xiong L., Stevenson B. & Zhu J.-K. 1997. Genetic analysis of osmotic and cold stress signal transduction in Arabidopsis: interactions and convergence of abscissic acid-dependent and abscissic acid-independent pathways. Plant Cell 9, 1935–1949.
Itano N. & Hatano S. 1991. F-actin binding protein from Physarum polycephalum: purification and its capacity for co-bundling of actin filaments and microtubules. Cell Motil & Cytoskeleton 19: 244–254.
Kandasamy M..K. & Meagher R.B. 1999. Actinorganelle interaction: association with chloroplasts in Arabidopsis leaf mesophyll cells. Cell Motil & Cytoskeleton 44, 110–118.
Laporte K., Rossignol M. & Traas J.A. 1993. Interaction of tubulin with the plasma membrane: tubulin is present in purified plasmalemma and behaves as an integral membrane protein. Planta 191, 413–416.
Lekka M., Lekki J., Schoulyarenko A.P., Cleff B. & Stachura J. 1996. Scanning force microscopy of biological samples. Polish Journal of Pathology 47, 51–55.
Lekka M., Laidler. P, Gil D., Lekki. J, Stachura Z. & Hrynkiewicz A.Z. 1999a. Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy. Eur Biophys J 28, 312–316.
Lekka M., Lekki J., Marszałek M., Golonka P., Stachura Z., Cleff B. & Hrynkiewicz A.Z. 1996b. Local elastic properties of cells studied by SFM. Appl Surf Sci 141, 345–349.
Lodish H., Baltimore D., Berk A., Zipursky S.L., Matsudaira P. & Darnell J. 1995. Molecular Cell Biology. Scientific American Books, An Imprint of WH Freeman and Company, New York, p 1061, p 1066.
Malec P., Rinaldi R.A. & Gabryś H. 1996. Light-induced chloroplast movements in Lemna trisulca. Identification of the motile system. Plant Sci 120, 127–137.
Masuda Y., Takagi S. & Nagai R. 1991. Proteasesensitive anchoring of microfilament bundles provides tracks for cytoplasmic streaming in Vallisneria. Protoplasma 162, 151–159.
Potrykus I. & Shillito R.D. 1986. Protoplasts: isolation, culture, plant regeneration. In: Methods in Enzymology Vol 118 (eds A. Weissbach & H. Weissbach), pp. 549–578. Academic Press.
Rezau C. & Pont-Lezica R.F. 1995. Comparing plant and animal extracellular matrix-cytoskeleton connections — are they alike? Protoplasma 186, 113–121.
Ryu J.-H., Mizuno K., Takagi S. & Nagai R. 1997. Extracellular components implicated in the stationary organisation of the actin cytoskeleton in mesophyll cells of Vallisneria. Plant Cell Physiol 38, 420–432.
Tao W. and Verbelen J.-P. 1996. Switching on and off cell expansion in cultured mesophyll protoplasts of tobacco. Plant Sci 116, 107–115.
Tominaga M., Morita K., Sonobe S., Yokota E. & Shimmen T. 1997. Microtubules regulate the organisation of actin filaments at the cortical region in root hair cells of Hydrocharis. Protoplasma 199, 83–92.
Trojan A. & Gabryś H. 1996. Chloroplast distribution in Arabidopsis thaliana (L.) depends on light conditions during growth. Plant Physiol 111, 419–425.
Wagner V.T., Brian L. & Quatrano R.S. 1992. Role of vitronectin-like molecule in embryo adhesion of the brown alga Fucus. Proc Natl Acad Sci USA 89, 3644–3648.
Walczak T., Gabryś H 1980. New type of photometer for measurements of transmission changes corresponding to chloroplast movements in leaves. Photosynthetica 14, 65–72.
Weisenhorn A., Khorsandi M., Kasas S., Gotzos V. & Butt H.J. 1993. Deformation and height anomaly of soft surfaces studied with an AFM. Nanotechnology 4, 106–113.
Wyatt S.E. & Carpita N.C. 1993. The plant cytoskeleton-cell-wall continuum. Trends Cell Biol 3, 413–417.
Yamada K.M. & Geiger B. 1997. Molecular interactions in cell adhesion complexes. Curr Opin Cell Biol 9, 76–85.
Zhu J.-K., Shi J., Singh U., Wyatt S.E., Bressan R.A., Hasegawa P.M. & Carpita N.C. 1993. Enrichment of vitronectin-and fibronectin-like proteins in NaCl-adapted plant cells and evidence for their involvement in plasma membrane-cell wall adhesion. Plant J 3, 637–646.
Zhu J.-K., Damasz B., Kononowicz A., Bressan R.A. & Hasegawa P.M. 1994. A higher plant extracellular vitronectin-like adhesion protein is related to the translational elongation factor-1α. Plant Cell 6, 393–404.
Zurzycki J. 1980. Blue-light induced intracellular movements. In: The Blue Light Syndrome (ed H. Senger), pp. 50–68. Springer, Berlin.
Zurzycki J., Walczak T., Gabryś H. & Kajfosz J. 1983. Chloroplast translocations in Lemna trisulca L. induced by continuous irradiation and by light pulses. Planta 157, 502–505.
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Augustynowicz, J., Lekka, M., Burda, K. et al. Correlation between chloroplast motility and elastic properties of tobacco mesophyll protoplasts. Acta Physiol Plant 23, 291–302 (2001). https://doi.org/10.1007/s11738-001-0036-7
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DOI: https://doi.org/10.1007/s11738-001-0036-7